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Please include the following statement when referencing the CPTAC Assay Portal
We would like to acknowledge the National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (CPTAC) Assay Portal (assays.cancer.gov) for developing assays and establishing criteria for the assays described in this publication.

Overview Data source: UniProt

Official Gene Symbol Other Aliases
MTOR FRAP, FRAP1, FRAP2, RAFT1, RAPT1
Sequence Length (AA) Molecular Weight (Da)
2549 288892
Protein Name
Serine/threonine-protein kinase mTOR
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MLGTGPAAAT TAATTSSNVS VLQQFASGLK SRNEETRAKA AKELQHYVTM
60 70 80 90 100
ELREMSQEES TRFYDQLNHH IFELVSSSDA NERKGGILAI ASLIGVEGGN
110 120 130 140 150
ATRIGRFANY LRNLLPSNDP VVMEMASKAI GRLAMAGDTF TAEYVEFEVK
160 170 180 190 200
RALEWLGADR NEGRRHAAVL VLRELAISVP TFFFQQVQPF FDNIFVAVWD
210 220 230 240 250
PKQAIREGAV AALRACLILT TQREPKEMQK PQWYRHTFEE AEKGFDETLA
260 270 280 290 300
KEKGMNRDDR IHGALLILNE LVRISSMEGE RLREEMEEIT QQQLVHDKYC
310 320 330 340 350
KDLMGFGTKP RHITPFTSFQ AVQPQQSNAL VGLLGYSSHQ GLMGFGTSPS
360 370 380 390 400
PAKSTLVESR CCRDLMEEKF DQVCQWVLKC RNSKNSLIQM TILNLLPRLA
410 420 430 440 450
AFRPSAFTDT QYLQDTMNHV LSCVKKEKER TAAFQALGLL SVAVRSEFKV
460 470 480 490 500
YLPRVLDIIR AALPPKDFAH KRQKAMQVDA TVFTCISMLA RAMGPGIQQD
510 520 530 540 550
IKELLEPMLA VGLSPALTAV LYDLSRQIPQ LKKDIQDGLL KMLSLVLMHK
560 570 580 590 600
PLRHPGMPKG LAHQLASPGL TTLPEASDVG SITLALRTLG SFEFEGHSLT
610 620 630 640 650
QFVRHCADHF LNSEHKEIRM EAARTCSRLL TPSIHLISGH AHVVSQTAVQ
660 670 680 690 700
VVADVLSKLL VVGITDPDPD IRYCVLASLD ERFDAHLAQA ENLQALFVAL
710 720 730 740 750
NDQVFEIREL AICTVGRLSS MNPAFVMPFL RKMLIQILTE LEHSGIGRIK
760 770 780 790 800
EQSARMLGHL VSNAPRLIRP YMEPILKALI LKLKDPDPDP NPGVINNVLA
810 820 830 840 850
TIGELAQVSG LEMRKWVDEL FIIIMDMLQD SSLLAKRQVA LWTLGQLVAS
860 870 880 890 900
TGYVVEPYRK YPTLLEVLLN FLKTEQNQGT RREAIRVLGL LGALDPYKHK
910 920 930 940 950
VNIGMIDQSR DASAVSLSES KSSQDSSDYS TSEMLVNMGN LPLDEFYPAV
960 970 980 990 1000
SMVALMRIFR DQSLSHHHTM VVQAITFIFK SLGLKCVQFL PQVMPTFLNV
1010 1020 1030 1040 1050
IRVCDGAIRE FLFQQLGMLV SFVKSHIRPY MDEIVTLMRE FWVMNTSIQS
1060 1070 1080 1090 1100
TIILLIEQIV VALGGEFKLY LPQLIPHMLR VFMHDNSPGR IVSIKLLAAI
1110 1120 1130 1140 1150
QLFGANLDDY LHLLLPPIVK LFDAPEAPLP SRKAALETVD RLTESLDFTD
1160 1170 1180 1190 1200
YASRIIHPIV RTLDQSPELR STAMDTLSSL VFQLGKKYQI FIPMVNKVLV
1210 1220 1230 1240 1250
RHRINHQRYD VLICRIVKGY TLADEEEDPL IYQHRMLRSG QGDALASGPV
1260 1270 1280 1290 1300
ETGPMKKLHV STINLQKAWG AARRVSKDDW LEWLRRLSLE LLKDSSSPSL
1310 1320 1330 1340 1350
RSCWALAQAY NPMARDLFNA AFVSCWSELN EDQQDELIRS IELALTSQDI
1360 1370 1380 1390 1400
AEVTQTLLNL AEFMEHSDKG PLPLRDDNGI VLLGERAAKC RAYAKALHYK
1410 1420 1430 1440 1450
ELEFQKGPTP AILESLISIN NKLQQPEAAA GVLEYAMKHF GELEIQATWY
1460 1470 1480 1490 1500
EKLHEWEDAL VAYDKKMDTN KDDPELMLGR MRCLEALGEW GQLHQQCCEK
1510 1520 1530 1540 1550
WTLVNDETQA KMARMAAAAA WGLGQWDSME EYTCMIPRDT HDGAFYRAVL
1560 1570 1580 1590 1600
ALHQDLFSLA QQCIDKARDL LDAELTAMAG ESYSRAYGAM VSCHMLSELE
1610 1620 1630 1640 1650
EVIQYKLVPE RREIIRQIWW ERLQGCQRIV EDWQKILMVR SLVVSPHEDM
1660 1670 1680 1690 1700
RTWLKYASLC GKSGRLALAH KTLVLLLGVD PSRQLDHPLP TVHPQVTYAY
1710 1720 1730 1740 1750
MKNMWKSARK IDAFQHMQHF VQTMQQQAQH AIATEDQQHK QELHKLMARC
1760 1770 1780 1790 1800
FLKLGEWQLN LQGINESTIP KVLQYYSAAT EHDRSWYKAW HAWAVMNFEA
1810 1820 1830 1840 1850
VLHYKHQNQA RDEKKKLRHA SGANITNATT AATTAATATT TASTEGSNSE
1860 1870 1880 1890 1900
SEAESTENSP TPSPLQKKVT EDLSKTLLMY TVPAVQGFFR SISLSRGNNL
1910 1920 1930 1940 1950
QDTLRVLTLW FDYGHWPDVN EALVEGVKAI QIDTWLQVIP QLIARIDTPR
1960 1970 1980 1990 2000
PLVGRLIHQL LTDIGRYHPQ ALIYPLTVAS KSTTTARHNA ANKILKNMCE
2010 2020 2030 2040 2050
HSNTLVQQAM MVSEELIRVA ILWHEMWHEG LEEASRLYFG ERNVKGMFEV
2060 2070 2080 2090 2100
LEPLHAMMER GPQTLKETSF NQAYGRDLME AQEWCRKYMK SGNVKDLTQA
2110 2120 2130 2140 2150
WDLYYHVFRR ISKQLPQLTS LELQYVSPKL LMCRDLELAV PGTYDPNQPI
2160 2170 2180 2190 2200
IRIQSIAPSL QVITSKQRPR KLTLMGSNGH EFVFLLKGHE DLRQDERVMQ
2210 2220 2230 2240 2250
LFGLVNTLLA NDPTSLRKNL SIQRYAVIPL STNSGLIGWV PHCDTLHALI
2260 2270 2280 2290 2300
RDYREKKKIL LNIEHRIMLR MAPDYDHLTL MQKVEVFEHA VNNTAGDDLA
2310 2320 2330 2340 2350
KLLWLKSPSS EVWFDRRTNY TRSLAVMSMV GYILGLGDRH PSNLMLDRLS
2360 2370 2380 2390 2400
GKILHIDFGD CFEVAMTREK FPEKIPFRLT RMLTNAMEVT GLDGNYRITC
2410 2420 2430 2440 2450
HTVMEVLREH KDSVMAVLEA FVYDPLLNWR LMDTNTKGNK RSRTRTDSYS
2460 2470 2480 2490 2500
AGQSVEILDG VELGEPAHKK TGTTVPESIH SFIGDGLVKP EALNKKAIQI
2510 2520 2530 2540 2549
INRVRDKLTG RDFSHDDTLD VPTQVELLIK QATSHENLCQ CYIGWCPFW

Data source: UniProt


Position of Targeted Peptide Analytes Relative to SNPs, Isoforms, and PTMs

Uniprot Database Entry PhosphoSitePlus ®

Click a point on a node
to view detailed assay information below
All other points link out to UniProt



Phosphorylation Acetylation Ubiquitylation Other

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Loader

Assay Details for CPTAC-1360 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
KLHVS[+80.0]TINLQK
Modification Type
Phospho (ST)
Protein - Site of Modification
1261
Peptide - Site of Modification
5
Peptide Start
1257
Peptide End
1267
CPTAC ID
CPTAC-1360
Peptide Molecular Mass
1,359.7276
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
digested cell lysate
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Mol Cell Proteomics. 2016 May;15(5):1622-41. doi: 10.1074/mcp.M116.058354. Epub 2016 Feb 24. Reduced-representation Phosphosignatures Measured by Quantitative Targeted MS Capture Cellular States and Enable Large-scale Comparison of Drug-induced Phenotypes. Abelin JG1, Patel J1, Lu X1, Feeney CM1, Fagbami L1, Creech AL1, Hu R1, Lam D1, Davison D1, Pino L1, Qiao JW1, Kuhn E1, Officer A1, Li J2, Abbatiello S1, Subramanian A1, Sidman R2, Snyder E3, Carr SA1, Jaffe JD4.

View Details (opens in a new window)

Mol Cell Proteomics. 2014 Jul;13(7):1690-704. doi: 10.1074/mcp.M113.036392. Epub 2014 Apr 9. Ischemia in tumors induces early and sustained phosphorylation changes in stress kinase pathways but does not affect global protein levels. Mertins P1, Yang F2, Liu T2, Mani DR3, Petyuk VA2, Gillette MA3, Clauser KR3, Qiao JW3, Gritsenko MA2, Moore RJ2, Levine DA4, Townsend R5, Erdmann-Gilmore P5, Snider JE5, Davies SR5, Ruggles KV6, Fenyo D6, Kitchens RT5, Li S5, Olvera N4, Dao F4, Rodriguez H7, Chan DW8, Liebler D9, White F10, Rodland KD2, Mills GB11, Smith RD2, Paulovich AG12, Ellis M5, Carr SA1


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva
Internal Standard
light stable isotope peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y9-98 (2+) 6.4 4.7 5.3 7 5.5 7 9.5 7.2 8.8 15 15 15
y9 (2+) 5 4.8 5.8 6.9 4.6 8.6 8.5 6.6 10.4 15 15 15
y8-98 (1+) 5.6 5.8 7.8 6.8 7.3 8.4 8.8 9.3 11.5 15 15 15
sum 5.3 4.5 5.3 5.7 5 7.3 7.8 6.7 9 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1361 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
LHVS[+80.0]TINLQK
Modification Type
Phospho (ST)
Protein - Site of Modification
1261
Peptide - Site of Modification
4
Peptide Start
1258
Peptide End
1267
CPTAC ID
CPTAC-1361
Peptide Molecular Mass
1,231.6326
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
digested cell lysate
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Mol Cell Proteomics. 2016 May;15(5):1622-41. doi: 10.1074/mcp.M116.058354. Epub 2016 Feb 24. Reduced-representation Phosphosignatures Measured by Quantitative Targeted MS Capture Cellular States and Enable Large-scale Comparison of Drug-induced Phenotypes. Abelin JG1, Patel J1, Lu X1, Feeney CM1, Fagbami L1, Creech AL1, Hu R1, Lam D1, Davison D1, Pino L1, Qiao JW1, Kuhn E1, Officer A1, Li J2, Abbatiello S1, Subramanian A1, Sidman R2, Snyder E3, Carr SA1, Jaffe JD4.

View Details (opens in a new window)

Mol Cell Proteomics. 2014 Jul;13(7):1690-704. doi: 10.1074/mcp.M113.036392. Epub 2014 Apr 9. Ischemia in tumors induces early and sustained phosphorylation changes in stress kinase pathways but does not affect global protein levels. Mertins P1, Yang F2, Liu T2, Mani DR3, Petyuk VA2, Gillette MA3, Clauser KR3, Qiao JW3, Gritsenko MA2, Moore RJ2, Levine DA4, Townsend R5, Erdmann-Gilmore P5, Snider JE5, Davies SR5, Ruggles KV6, Fenyo D6, Kitchens RT5, Li S5, Olvera N4, Dao F4, Rodriguez H7, Chan DW8, Liebler D9, White F10, Rodland KD2, Mills GB11, Smith RD2, Paulovich AG12, Ellis M5, Carr SA1


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva
Internal Standard
light stable isotope peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y4 (1+) 4 4 4.7 5.2 10.2 10.3 6.6 11 11.3 15 15 15
y6 (1+) 5.1 5 4.6 6.1 6 10 8 7.8 11 15 15 15
y8-98 (1+) 3.4 3.5 2.5 4.6 7.3 8.2 5.7 8.1 8.6 15 15 15
sum 3.5 3.6 3.1 3.7 7 8.8 5.1 7.9 9.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5753 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
DLELAVPGTYDPNQPIIR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2135
Peptide End
2152
CPTAC ID
CPTAC-5753
Peptide Molecular Mass
2,010.0422
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y12 (1+) 57.7 17.3 13.6 56.8 37.3 25.5 81 41.1 28.9 15 15 15
y7 (1+) 38.7 16.6 13.7 37.9 38.1 22.9 54.2 41.6 26.7 15 15 15
b5 (1+) 51.3 16.1 13.3 47.2 40.6 25.3 69.7 43.7 28.6 15 15 15
sum 44.2 15.3 13.5 41.3 37.7 24.7 60.5 40.7 28.1 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5752 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
IQSIAPSLQVITSK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2153
Peptide End
2166
CPTAC ID
CPTAC-5752
Peptide Molecular Mass
1,483.8610
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y10 (1+) 21.1 11.8 7.9 28.7 32.5 18.4 35.6 34.6 20 15 15 15
y9 (1+) 23.3 10.3 6.9 26.4 32.9 17.8 35.2 34.5 19.1 15 15 15
sum 21.8 10.7 7.1 26.3 32.7 18 34.2 34.4 19.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5755 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
LFDAPEAPLPSR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
1121
Peptide End
1132
CPTAC ID
CPTAC-5755
Peptide Molecular Mass
1,311.6823
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y10 (1+) 24.9 10.1 7.4 29.2 23.4 8.9 38.4 25.5 11.6 15 15 15
y8 (1+) 22.2 8.8 7.6 24.2 23.7 9.8 32.8 25.3 12.4 15 15 15
sum 21.3 9.1 7.5 24.5 23.6 9.6 32.5 25.3 12.2 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5754 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
LTESLDFTDYASR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
1142
Peptide End
1154
CPTAC ID
CPTAC-5754
Peptide Molecular Mass
1,516.7046
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y8 (1+) 17.9 16 11.6 21.7 33.4 16.2 28.1 37 19.9 15 15 15
y7 (1+) 29.4 16.9 11.5 33.7 33.1 16.8 44.7 37.2 20.4 15 15 15
y6 (1+) 23.5 15.1 10.8 26.1 33.6 15.9 35.1 36.8 19.2 15 15 15
y4 (1+) 29.7 15.3 11.3 34.3 34.4 16.5 45.4 37.6 20 15 15 15
sum 23 15.5 11.3 26.5 33.5 16.2 35.1 36.9 19.8 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5798 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TDS[+79.966331]YSAGQSVEILDGVELGEPAHK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2446
Peptide End
2469
CPTAC ID
CPTAC-5798
Peptide Molecular Mass
2,581.1585
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y6 (1+) 22.5 12.8 13.5 23.9 15.5 13.8 32.8 20.1 19.3 16 15 16
y8 (1+) 19 11.8 16.7 20 13.8 15 27.6 18.2 22.4 16 15 16
sum 20.2 12.2 12.2 22.2 14.5 11.6 30 18.9 16.8 16 15 16


Additional Resources and Comments


Assay Details for CPTAC-5797 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TDSYSAGQSVEILDGVELGEPAHK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2446
Peptide End
2469
CPTAC ID
CPTAC-5797
Peptide Molecular Mass
2,501.1922
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y6 (1+) 34.6 13.6 8.5 44.4 15.7 10.6 56.3 20.8 13.6 16 15 17
y4 (1+) 36.5 9.4 8.9 37.9 16.9 15.3 52.6 19.3 17.7 16 15 17
b3 (1+) 27.8 11.2 8 22.9 13.3 9.5 36 17.4 12.4 16 15 17
sum 25.6 9 4.8 25.3 12.9 9.8 36 15.7 10.9 16 15 17


Additional Resources and Comments


Assay Details for CPTAC-5794 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TGTTVPESIHSFIGDGLVKPEALNK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2471
Peptide End
2495
CPTAC ID
CPTAC-5794
Peptide Molecular Mass
2,609.3701
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y6 (1+) 15.7 8 21 16.5 9.2 21.4 22.8 12.2 30 16 15 17
y20 (3+) 13.2 5.9 19.1 10.8 6.6 21.4 17.1 8.9 28.7 16 15 17
sum 13 5.9 18.9 10.6 6.5 21.3 16.8 8.8 28.5 16 15 17


Additional Resources and Comments


Assay Details for CPTAC-5793 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TGTTVPESIHS[+79.966331]FIGDGLVKPEALNK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2471
Peptide End
2495
CPTAC ID
CPTAC-5793
Peptide Molecular Mass
2,689.3364
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y19 (3+) 32.1 13.7 39.1 29.4 24.6 41.2 43.5 28.2 56.8 16 15 16
y20 (3+) 15.2 10 37.5 24.1 19.7 41.5 28.5 22.1 55.9 16 15 16
sum 11.7 9.4 37.5 19.3 20.3 41.4 22.6 22.4 55.9 16 15 16


Additional Resources and Comments


Assay Details for CPTAC-5796 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TGTTVPESIHSFIGDGLVKPEALNKK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2471
Peptide End
2496
CPTAC ID
CPTAC-5796
Peptide Molecular Mass
2,737.4650
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y7 (1+) 51.2 22.1 13.4 54.2 22.9 17.9 74.6 31.8 22.4 16 15 17
y21 (2+) 23.7 13.1 7.9 22.3 18.5 7.2 32.5 22.7 10.7 16 15 17
b5 (1+) 47.9 18.9 10.2 44.6 17.5 11.9 65.4 25.8 15.7 16 15 17
sum 19.9 12 7.2 20.5 16.5 6.9 28.6 20.4 10 16 15 17


Additional Resources and Comments


Assay Details for CPTAC-5795 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TGTTVPESIHS[+79.966331]FIGDGLVKPEALNKK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2471
Peptide End
2496
CPTAC ID
CPTAC-5795
Peptide Molecular Mass
2,817.4314
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y22 (2+) 27.9 13.5 28.2 27.3 17.9 27.4 39 22.4 39.3 16 15 16
b5 (1+) 14.8 15.5 29.4 13.5 17.2 28.5 20 23.2 40.9 16 15 16
y7 (1+) 15.3 13.4 25.7 22.9 14.6 26.4 27.5 19.8 36.8 16 15 16
sum 14.3 12.8 26.4 14.9 14.6 26.4 20.7 19.4 37.3 16 15 16


Additional Resources and Comments


Assay Details for CPTAC-5800 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TRTDS[+79.966331]YSAGQSVEILDGVELGEPAHK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2444
Peptide End
2469
CPTAC ID
CPTAC-5800
Peptide Molecular Mass
2,838.3073
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y4 (1+) 14.3 11.6 9.8 16.2 13.1 10 21.6 17.5 14 16 15 16
y6 (1+) 24 15 7.8 27.8 17.3 8.3 36.7 22.9 11.4 16 15 16
y11 (1+) 18.2 11.8 17 17 16.1 20.7 24.9 20 26.8 16 15 16
sum 17.2 12.5 8.9 18.6 15 9.8 25.3 19.5 13.2 16 15 16


Additional Resources and Comments


Assay Details for CPTAC-5799 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TRTDSYSAGQSVEILDGVELGEPAHK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2444
Peptide End
2469
CPTAC ID
CPTAC-5799
Peptide Molecular Mass
2,758.3410
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y11 (1+) 55.6 19.8 15.2 60.8 27.2 17.6 82.4 33.6 23.3 16 15 17
y6 (1+) 27.5 11.5 15.8 37.2 34.3 19.4 46.3 36.2 25 16 15 17
y4 (1+) 48.4 16.3 17.1 54.8 22.9 17.7 73.1 28.1 24.6 16 15 17
sum 30.2 10.1 13.4 36.1 22.8 14.9 47.1 24.9 20 16 15 17


Additional Resources and Comments


Assay Details for CPTAC-5756 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
VLGLLGALDPYK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
887
Peptide End
898
CPTAC ID
CPTAC-5756
Peptide Molecular Mass
1,257.7333
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y10 (1+) 38 20.4 16.7 33.9 45.3 28.3 50.9 49.7 32.9 15 15 15
y8 (1+) 48.2 18.8 16.6 43.6 46 29.5 65 49.7 33.8 15 15 15
y7 (1+) 42.2 18.2 16.1 36.4 44.6 29 55.7 48.2 33.2 15 15 15
sum 41.6 18.8 16.3 36 45.1 28.9 55 48.9 33.2 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-6111 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
LFDAPEAPLPSR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
1121
Peptide End
1132
CPTAC ID
non-CPTAC-6111
Peptide Molecular Mass
1,311.6823
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Bovine Serum Albumin
Submitting Laboratory
McGill University
Submitting Lab PI
Christoph H. Borchers, Gerald Batist

Publication

View Details (opens in a new window)

mTORC1-driven protein translation correlates with clinical benefit of capivasertib within a genetically preselected cohort of PIK3CA-altered tumours. Constance A Sobsey, Bjoern C Froehlich, Georgia Mitsa, Sahar Ibrahim, Robert Popp, Rene P Zahedi, Elza C de Bruin, Christoph H Borchers, Gerald Batist


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Agilent 6495B QQQ-MS
Internal Standard
Standard Isotope labeled internal standard peptides with 13Cx, 15Ny (R+10 or K+8)
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Infinity 1290
Column Packing
commercial (Agilent ZORBAX Eclipse Plus C18)
Column Dimensions
4.6 mm × 150 mm, (5 µm)
Flow Rate
0.4 mL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
b3 (1+) 17 8.9 6.8 16.6 10.6 5.7 23.8 13.8 8.9 16 16 17
y10 (1+) 23.1 9.9 6.8 19.6 9.2 7.8 30.3 13.5 10.3 16 16 17
y5 (1+) 15.1 9.5 6.4 17.2 9.5 4.8 22.9 13.4 8 16 16 17
y8 (1+) 17.4 7.7 8.4 18.6 6 8.9 25.5 9.8 12.2 16 16 17
y8 (2+) 34.5 8.8 8.2 42.7 6.7 6.2 54.9 11.1 10.3 16 16 17
sum 7.4 3.4 2.9 7.7 3.4 4 10.7 4.8 4.9 16 16 17


Additional Resources and Comments


Assay Details for non-CPTAC-5563 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
DLELAVPGTYDPNQPIIR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
6
Peptide Start
2135
Peptide End
2152
CPTAC ID
non-CPTAC-5563
Peptide Molecular Mass
2,010.0422
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Line
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
SIS Peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
RSLCnano
Column Packing
C18 PepMap100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
b6 (1+) 48.6 13.5 12 45.5 40.5 23.1 66.6 42.7 26 14 14 14
b5 (1+) 40.6 12.9 12.5 33.5 41.9 24 52.6 43.8 27.1 14 14 14
y4 (1+) 77.5 27.8 14.2 98.6 34.4 22.1 125.4 44.2 26.3 14 14 14
y7 (1+) 32.3 12.4 12.6 35.5 32.9 22.3 48 35.2 25.6 14 14 14
y12 (1+) 45.5 17.4 11.5 49.7 38.8 22.7 67.4 42.5 25.4 14 14 14
sum 38.1 13.5 12 36.5 38.4 22.9 52.8 40.7 25.9 14 14 14


Additional Resources and Comments


Assay Details for non-CPTAC-5421 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
IQSIAPSLQVITSK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
2153
Peptide End
2166
CPTAC ID
non-CPTAC-5421
Peptide Molecular Mass
1,483.8610
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y10 (1+) 9.3 4.4 2.6 9 4.9 3.3 12.9 6.6 4.2 15 15 15
y11 (1+) 10.4 8.4 6 10.6 8.7 7.2 14.8 12.1 9.4 15 15 15
y12 (1+) 6.6 4.4 2.2 6.3 4.6 3.8 9.1 6.4 4.4 15 15 15
y9 (1+) 4.9 4.7 2.5 5.2 5.3 2.9 7.1 7.1 3.8 15 15 15
sum 3.4 3.9 1.5 3.8 4.7 2.8 5.1 6.1 3.2 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5561 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
LFDAPEAPLPSR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
6
Peptide Start
1121
Peptide End
1132
CPTAC ID
non-CPTAC-5561
Peptide Molecular Mass
1,311.6823
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Line
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
SIS Peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
RSLCnano
Column Packing
C18 PepMap100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
b3 (1+) 22 7.4 7.9 22.4 23.9 9 31.4 25 12 14 14 14
y5 (1+) 20 6.3 7.7 21.3 23.3 9.5 29.2 24.1 12.2 14 14 14
y8 (1+) 21.7 7.1 8.1 26.3 24.6 9.4 34.1 25.6 12.4 14 14 14
y10 (1+) 24.2 7 8.3 28.6 24.1 9.2 37.5 25.1 12.4 14 14 14
sum 21.4 6.8 7.9 24 24.1 9.2 32.2 25 12.1 14 14 14


Additional Resources and Comments


Assay Details for non-CPTAC-5562 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
LTESLDFTDYASR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
6
Peptide Start
1142
Peptide End
1154
CPTAC ID
non-CPTAC-5562
Peptide Molecular Mass
1,516.7046
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Line
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
SIS Peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
RSLCnano
Column Packing
C18 PepMap100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y4 (1+) 23.2 14.2 12 27.5 34.1 15 36 36.9 19.2 14 14 14
y6 (1+) 22.1 12.3 11.9 26 32.6 16.1 34.1 34.8 20 14 14 14
y7 (1+) 23.1 13.4 12.4 29 32 15.9 37.1 34.7 20.2 14 14 14
y8 (1+) 19 14 12.5 22.9 33.1 15.4 29.8 35.9 19.8 14 14 14
y10 (1+) 16.3 13.5 12.6 25.8 33.1 14.5 30.5 35.7 19.2 14 14 14
sum 20.2 13.3 12.2 24.9 32.9 15.4 32.1 35.5 19.6 14 14 14


Additional Resources and Comments


Assay Details for non-CPTAC-5711 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TDS[+79.966331]YSAGQSVEILDGVELGEPAHK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2446
Peptide End
2469
CPTAC ID
non-CPTAC-5711
Peptide Molecular Mass
2,581.1585
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y11 (1+) 32.3 8.3 5 18.3 10.1 5.5 37.1 13.1 7.4 15 15 15
y10 (1+) 24.3 9.3 6.5 10 9.5 8.4 26.3 13.3 10.6 15 15 15
y8 (1+) 25.8 8.8 6.8 6.3 6.1 6.5 26.6 10.7 9.4 15 15 15
y6 (1+) 18.9 5.9 1.8 6.7 4.8 2.5 20.1 7.6 3.1 15 15 15
y4 (1+) 20.6 5.6 3 8.7 4.6 3.7 22.4 7.2 4.8 15 15 15
sum 21.2 5.5 2.7 4.7 4.3 3.4 21.7 7 4.3 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5423 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TDSYSAGQSVEILDGVELGEPAHK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
2446
Peptide End
2469
CPTAC ID
non-CPTAC-5423
Peptide Molecular Mass
2,501.1922
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y18 (2+) 12.4 6.7 3.4 12.3 7 4 17.5 9.7 5.2 15 15 15
y19 (2+) 20.4 6.8 6.7 20.3 8.3 8.4 28.8 10.7 10.7 15 15 15
y20 (2+) 4.1 4.9 4.7 4.6 6.7 4.9 6.2 8.3 6.8 15 15 15
y4 (1+) 10.9 4.1 3.6 13.9 4.7 3.8 17.7 6.2 5.2 15 15 15
y6 (1+) 15.8 5.1 9.4 14.6 5.3 9.2 21.5 7.4 13.2 15 15 15
sum 5.6 2.5 2.4 5.8 4.1 3.2 8.1 4.8 4 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5709 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TGTTVPESIHS[+79.966331]FIGDGLVKPEALNK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2471
Peptide End
2495
CPTAC ID
non-CPTAC-5709
Peptide Molecular Mass
2,689.3364
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y7 (1+) 8.3 3.7 4.6 9 4.8 5.3 12.2 6.1 7 15 15 15
y6 (1+) 3 2.5 2.1 3.2 2.3 2.4 4.4 3.4 3.2 15 15 15
y20-98 (2+) 3.9 2.5 3.4 3.9 2.7 4.4 5.5 3.7 5.6 15 15 15
y19 (2+) 9.6 6.3 2.8 10.3 5.7 2.7 14.1 8.5 3.9 15 15 15
b11 (3+) 7.2 6.5 2 6.9 6.5 3.7 10 9.2 4.2 15 15 15
sum 2.7 2.9 1.6 2.6 2.6 2.9 3.7 3.9 3.3 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5424 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TGTTVPESIHSFIGDGLVKPEALNK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
2471
Peptide End
2495
CPTAC ID
non-CPTAC-5424
Peptide Molecular Mass
2,609.3701
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y20 (3+) 4.1 3.9 1.7 4.2 6 3.7 5.9 7.2 4.1 15 15 15
y21 (3+) 13.2 7.5 4.1 12.5 8.3 5.1 18.2 11.2 6.5 15 15 15
y2 (1+) 20.7 6.6 4.9 24.4 7.8 5.3 32 10.2 7.2 15 15 15
y6 (1+) 10.2 5.1 4.4 10.3 6.7 4.9 14.5 8.4 6.6 15 15 15
sum 3.1 4 1.7 3.4 6.1 3.6 4.6 7.3 4 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5710 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TGTTVPESIHS[+79.966331]FIGDGLVKPEALNKK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2471
Peptide End
2496
CPTAC ID
non-CPTAC-5710
Peptide Molecular Mass
2,817.4314
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y8 (1+) 26.8 8.7 11.9 24.3 12.6 12.5 36.2 15.3 17.3 15 15 15
y7 (1+) 7.8 4.4 5.2 5.6 4.9 5.1 9.6 6.6 7.3 15 15 15
y21-98 (2+) 5.3 5.8 5.9 6.9 6.7 6.6 8.7 8.9 8.9 15 15 15
y20 (2+) 13.6 6.3 6.5 11.3 4.6 8.8 17.7 7.8 10.9 15 15 15
b5 (1+) 14.2 4.9 5.3 14.3 5.7 6.4 20.2 7.5 8.3 15 15 15
sum 6 3.9 3.4 5.4 4.2 3.7 8.1 5.7 5 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5425 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Sequence
TGTTVPESIHSFIGDGLVKPEALNKK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
2471
Peptide End
2496
CPTAC ID
non-CPTAC-5425
Peptide Molecular Mass
2,737.4650
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

Loader

Repeatability

Data source: Panorama

  Average intra-assay CV
(within day CV)
Average inter-assay CV
(between day CV)
Total CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y21 (3+) 3.5 4.1 2.1 4.3 5.3 3.2 5.5 6.7 3.8 15 15 15
y22 (3+) 20.3 4.8 2.8 27.1 6.9 4.3 33.9 8.4 5.1 15 15 15
y23 (3+) 16.7 7.3 4.2 20.7 7.9 4.6 26.6 10.8 6.2 15 15 15
y21 (2+) 14.3 10.2 7.9 15.1 10.1 9.5 20.8 14.4 12.4 15 15 15
y7 (1+) 6.1 6.1 2.9 8.6 7.1 3.8 10.5 9.4 4.8 15 15 15
sum 2.2 3.5 1.6 3.6 5 2.9 4.2 6.1 3.3 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5712 Collapse assay details

Data source: Panorama

Official Gene Symbol
MTOR
Peptide Modified Sequence
TRTDS[+79.966331]YSAGQSVEILDGVELGEPAHK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
2444
Peptide End
2469
CPTAC ID
non-CPTAC-5712
Peptide Molecular Mass
2,838.3073
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Stress Pathway Panel


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama