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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
AKT1 PKB, RAC
Sequence Length (AA) Molecular Weight (Da)
480 55686
Protein Name
RAC-alpha serine/threonine-protein kinase
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MSDVAIVKEG WLHKRGEYIK TWRPRYFLLK NDGTFIGYKE RPQDVDQREA
60 70 80 90 100
PLNNFSVAQC QLMKTERPRP NTFIIRCLQW TTVIERTFHV ETPEEREEWT
110 120 130 140 150
TAIQTVADGL KKQEEEEMDF RSGSPSDNSG AEEMEVSLAK PKHRVTMNEF
160 170 180 190 200
EYLKLLGKGT FGKVILVKEK ATGRYYAMKI LKKEVIVAKD EVAHTLTENR
210 220 230 240 250
VLQNSRHPFL TALKYSFQTH DRLCFVMEYA NGGELFFHLS RERVFSEDRA
260 270 280 290 300
RFYGAEIVSA LDYLHSEKNV VYRDLKLENL MLDKDGHIKI TDFGLCKEGI
310 320 330 340 350
KDGATMKTFC GTPEYLAPEV LEDNDYGRAV DWWGLGVVMY EMMCGRLPFY
360 370 380 390 400
NQDHEKLFEL ILMEEIRFPR TLGPEAKSLL SGLLKKDPKQ RLGGGSEDAK
410 420 430 440 450
EIMQHRFFAG IVWQHVYEKK LSPPFKPQVT SETDTRYFDE EFTAQMITIT
460 470 480
PPDQDDSMEC VDSERRPHFP QFSYSASGTA

Data source: UniProt


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

Uniprot Database Entry PhosphoSitePlus ®

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to view detailed assay information below
All other points link out to UniProt



Phosphorylation Acetylation Ubiquitylation Other

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Assay Details for non-CPTAC-6248 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
RPHFPQFS[+79.966331]YSASGTA
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
8
Peptide Start
466
Peptide End
480
CPTAC ID
non-CPTAC-6248
Peptide Molecular Mass
1,731.7406
Species
Human
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
Matrix
SKBR3 cells
Submitting Laboratory
Cell Signaling Technology
Submitting Lab PI
Yiying Zhu

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Orbitrap Q Exactive (Thermo)
Internal Standard
Heavy isotope-labeled peptides
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N F
LC
EASY-nLC II (Thermo)
Column Packing
self-packed
Column Dimensions
ID 100um, length 15cm
Flow Rate
300 nL/min

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
b2 (1+) 8 6.3 7.8 10 13.4 10.3 12.8 14.8 12.9 15 15 15
b6 (1+) 15.1 6.8 4.9 17.6 12.4 7.3 23.2 14.1 8.8 15 15 15
b4 (1+) 5.8 6.4 7.3 7.3 12.4 10 9.3 14 12.4 15 15 15
b8-98 (2+) 6.9 7.2 5.4 11.7 15.4 8.4 13.6 17 10 15 15 15
b9-98 (2+) 7.6 5.6 6.2 11.4 13.2 10.3 13.7 14.3 12 15 15 15
sum 6.2 6 6.6 8.7 13.1 9.5 10.7 14.4 11.6 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5758 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FFAGIVWQHVYEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
407
Peptide End
419
CPTAC ID
CPTAC-5758
Peptide Molecular Mass
1,622.8245
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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

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+) 62 14 14.3 60 19.8 13.2 86.3 24.2 19.5 13 14 15
y5 (1+) 37.3 33.1 35.8 31.5 29.9 31.2 48.8 44.6 47.5 14 14 15
y6 (1+) 59.6 20.8 21.3 58.6 28.4 25.2 83.6 35.2 33 14 14 15
sum 45.9 21.8 23.2 40.5 20.8 15.8 61.2 30.1 28.1 14 14 15


Additional Resources and Comments


Assay Details for CPTAC-5806 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
RPHFPQFSYSASGTA
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
466
Peptide End
480
CPTAC ID
CPTAC-5806
Peptide Molecular Mass
1,651.7743
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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 F
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

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
b4 (1+) 5.3 5.3 10.9 8.9 16.9 18.2 10.4 17.7 21.2 16 15 17
b10 (2+) 11.1 6.9 8.5 12.3 17.8 16 16.6 19.1 18.1 16 15 17
sum 6 4.5 9.8 8.3 16.9 17.3 10.2 17.5 19.9 16 15 17


Additional Resources and Comments


Assay Details for CPTAC-5805 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
RPHFPQFS[+79.966331]YSASGTA
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
466
Peptide End
480
CPTAC ID
CPTAC-5805
Peptide Molecular Mass
1,731.7406
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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 F
LC
Dionex Ultimate 3000
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

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
b11 (2+) 20.8 17.1 19.5 25.5 22.3 29.7 32.9 28.1 35.5 16 15 16
b4 (1+) 8.7 12.1 15.4 19.3 22.5 21.5 21.2 25.5 26.4 16 15 16
b8 (2+) 21.4 18.2 12.1 20.1 21.7 23.7 29.4 28.3 26.6 16 15 16
sum 9.7 14 12.8 15.3 22.2 23.2 18.1 26.2 26.5 16 15 16


Additional Resources and Comments


Assay Details for CPTAC-5759 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
SLLSGLLK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
378
Peptide End
385
CPTAC ID
CPTAC-5759
Peptide Molecular Mass
829.5273
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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

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+) 36.1 11.8 13.3 45 25.6 24.1 57.7 28.2 27.5 15 15 15
y5 (1+) 19.7 13.1 8.2 38.8 35.7 18.2 43.5 38 20 15 15 15
y6 (1+) 22.7 12.1 8.9 24.7 34.3 18.7 33.5 36.4 20.7 15 15 15
sum 19.7 11.9 8.7 26.8 33.9 18.7 33.3 35.9 20.6 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5808 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
TFC[+57.021464]GTPEYLAPEVLEDNDYGR
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
308
Peptide End
328
CPTAC ID
CPTAC-5808
Peptide Molecular Mass
2,445.0795
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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

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+) 37.5 8.7 15.2 39 21.2 19.5 54.1 22.9 24.7 16 15 17
y7 (1+) 39.8 13.6 10.1 36.9 30 15.3 54.3 32.9 18.3 16 15 17
sum 26.5 7.8 11.7 29.5 25.5 16.7 39.7 26.7 20.4 16 15 17


Additional Resources and Comments


Assay Details for CPTAC-5807 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
T[+79.966331]FC[+57.021464]GTPEYLAPEVLEDNDYGR
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
308
Peptide End
328
CPTAC ID
CPTAC-5807
Peptide Molecular Mass
2,525.0458
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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

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+) 11.4 10.9 24.8 10.6 14 33.2 15.6 17.7 41.4 16 15 16
y11 (2+) 10.9 11.3 8.6 11.3 12.6 9.1 15.7 16.9 12.5 16 15 16
sum 11 11.1 8.6 10.9 12.9 10.1 15.5 17 13.3 16 15 16


Additional Resources and Comments


Assay Details for non-CPTAC-6077 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FFAGIVWQHVYEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
407
Peptide End
419
CPTAC ID
non-CPTAC-6077
Peptide Molecular Mass
1,622.8245
Species
Homo sapiens (Human)
Assay Type
Direct MRM-MS
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 K
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

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
b2 (1+) 16.5 4.4 6.2 25.4 3.7 9.7 30.3 5.7 11.5 15 16 17
y10 (2+) 20.2 6 6.5 25 6.2 6.7 32.1 8.6 9.3 15 16 17
y11 (2+) 15 4.4 6.5 19.4 6.7 7.2 24.5 8 9.7 15 16 17
y12 (2+) 23.5 12.6 8.4 30.9 12.7 6.5 38.8 17.9 10.6 15 16 17
y8 (2+) 22.2 5 9.1 22.6 4.5 9.2 31.7 6.7 12.9 15 16 17
sum 15.3 2.6 3.5 19.4 4.3 4.9 24.7 5 6 15 16 17


Stability

Data source: Panorama

Fragment ion / Transition control_intra_CV actual_temp_intra_CV frozen_intra_CV FTx1_intra_CV FTx2_intra_CV
b2 (1+) 7.5 2.9 2.7 6.3 2.6
y10 (2+) 4.8 6.5 7.7 3.5 2.2
y11 (2+) 7.1 7 4.2 3.2 5.7
y12 (2+) 13.1 11.9 6.8 8.3 6.2
y8 (2+) 7.4 9 3.5 13.9 6.3
sum 4.9 3.2 1.9 1.9 3
Fragment ion / Transition all_intra_CV all_inter_CV
b2 (1+) 4.1 5.2
y10 (2+) 5.2 5.2
y11 (2+) 5.7 4.7
y12 (2+) 9.7 9.2
y8 (2+) 8.2 8.9
sum 3 3.1

Endogenous

Data source: Panorama

Fragment ion / Transition intra_CV inter_CV total_CV
b2 (1+) 18.3 31.1 36.1
y10 (2+) 38.8 52.3 65.1
y11 (2+) 20.8 20.6 29.3
y12 (2+) 18.5 38.1 42.4
y8 (2+) 5.6 39.4 39.8
sum 18.2 20.5 27.4

Additional Resources and Comments


Assay Details for non-CPTAC-5528 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FFAGIVWQHVYEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
6
Peptide Start
407
Peptide End
419
CPTAC ID
non-CPTAC-5528
Peptide Molecular Mass
1,622.8245
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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 K
LC
RSLCnano
Column Packing
C18 PepMap100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

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
y5 (1+) 64.7 30.3 44 27.1 36.1 44.7 70.1 47.1 62.7 10 10 11
y7 (1+) 55.1 19.1 28.5 45.9 19.6 37.4 71.7 27.4 47 10 10 11
y8 (1+) 88.4 28.4 25.2 52.6 36.1 27.2 102.9 45.9 37.1 10 10 11
y10 (1+) 83.2 40.1 28.3 81.3 65.2 29.8 116.3 76.5 41.1 10 10 11
sum 59.3 24.2 31.6 27.2 27.9 36.7 65.2 36.9 48.4 10 10 11


Additional Resources and Comments


Assay Details for non-CPTAC-5328 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FYGAEIVSALDYLHSEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
252
Peptide End
268
CPTAC ID
non-CPTAC-5328
Peptide Molecular Mass
1,940.9520
Species
Homo sapiens (Human)
Assay Type
Direct MRM-MS
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

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+) 16.2 3.5 4.9 15.6 11.2 4.8 22.5 11.7 6.9 15 15 15
y10 (1+) 12.2 6.3 4 19.4 11.8 6.4 22.9 13.4 7.5 15 15 15
y11 (1+) 11 4.6 4.6 15 10.1 6.2 18.6 11.1 7.7 15 15 15
sum 8.2 4.2 2.9 14.2 10.8 4.4 16.4 11.6 5.3 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5332 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
RPHFPQFSYSASGTA
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
466
Peptide End
480
CPTAC ID
non-CPTAC-5332
Peptide Molecular Mass
1,651.7743
Species
Homo sapiens (Human)
Assay Type
Direct MRM-MS
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 F
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

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
b10 (2+) 20 9.4 10.1 19.6 13 11.3 28 16 15.2 15 15 15
b6 (1+) 34 5.5 5.7 36.5 10.3 5.2 49.9 11.7 7.7 15 15 15
b4 (1+) 12.7 5.9 3.2 16.3 5.3 3.9 20.7 7.9 5 15 15 15
sum 6.5 5.8 2.8 6.7 6.4 4.3 9.3 8.6 5.1 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5720 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
RPHFPQFS[+79.966331]YSASGTA
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
466
Peptide End
480
CPTAC ID
non-CPTAC-5720
Peptide Molecular Mass
1,731.7406
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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 F
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

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+) 13.2 7.9 5.3 17.5 13.2 4.5 21.9 15.4 7 13 12 13
b3 (1+) 21.4 14.9 19.2 33.9 24.9 18.2 40.1 29 26.5 13 12 13
b4 (1+) 9.4 7.4 5.7 11.1 7.4 7.5 14.5 10.5 9.4 13 12 13
sum 8.2 5.9 8.4 9.6 10.1 8.1 12.6 11.7 11.7 13 12 13


Additional Resources and Comments


Assay Details for non-CPTAC-5527 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
SLLSGLLK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
6
Peptide Start
378
Peptide End
385
CPTAC ID
non-CPTAC-5527
Peptide Molecular Mass
829.5273
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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 K
LC
RSLCnano
Column Packing
C18 PepMap100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nL/min

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+) 36.8 6.8 11.2 45.3 26.3 22.5 58.4 27.2 25.1 14 14 14
y5 (1+) 20.1 4.7 8.6 40.2 34.2 18.7 44.9 34.5 20.6 14 14 14
y6 (1+) 23.5 8.8 9.2 27.7 36.1 18.9 36.3 37.2 21 14 14 14
sum 21.2 7.3 9.1 29.9 35.1 19 36.7 35.9 21.1 14 14 14


Additional Resources and Comments


Assay Details for non-CPTAC-5719 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
T[+79.966331]FC[+57.021464]GTPEYLAPEVLEDNDYGR
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
308
Peptide End
328
CPTAC ID
non-CPTAC-5719
Peptide Molecular Mass
2,525.0458
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM-MS
Enrichment Method
N/A
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 R
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
Flow Rate
300 nL/min

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+) 7.2 4.9 3.9 4.6 4.5 3.5 8.5 6.7 5.2 15 15 15
y8 (1+) 5.3 2.3 3.4 6.1 3 3.1 8.1 3.8 4.6 15 15 15
y7 (1+) 5.7 2.7 2.1 5.7 2.3 2.1 8.1 3.5 3 15 15 15
y6 (1+) 5.2 4.4 2.4 4.6 5.2 2.6 6.9 6.8 3.5 15 15 15
y3 (1+) 3.9 4.3 2.5 3.7 5.2 3 5.4 6.7 3.9 15 15 15
sum 2.7 2.6 1.6 2.5 2.6 1.5 3.7 3.7 2.2 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5329 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
TFC[+57.021464]GTPEYLAPEVLEDNDYGR
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
311
Peptide - Site of Modification
3
Peptide Start
308
Peptide End
328
CPTAC ID
non-CPTAC-5329
Peptide Molecular Mass
2,445.0795
Species
Homo sapiens (Human)
Assay Type
Direct MRM-MS
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 R
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
Flow Rate
300 nl/min

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+) 16.1 4.6 4.3 14.9 8.4 8.3 21.9 9.6 9.3 15 15 15
y8 (1+) 13.7 7.4 4.7 14.5 11.4 5.2 19.9 13.6 7 15 15 15
y11 (1+) 17 6.5 4.6 18.8 6.4 5.9 25.3 9.1 7.5 15 15 15
sum 12.5 3.3 2 10 6.9 5.2 16 7.6 5.6 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3164 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
EVIVAKDEVAHTLTENR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
184
Peptide End
200
CPTAC ID
CPTAC-3164
Peptide Molecular Mass
1,923.0062
Species
Homo Sapiens
Assay Type
Direct MRM-MS
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

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+) 23.9 9.4 11.8 22.8 7.5 11.6 33 12 16.5 15 15 15
y5 (1+) 21.4 9.2 11.8 22.2 8.3 12.3 30.8 12.4 17 15 15 15
y7 (1+) 23 10.7 12.4 23.7 10 13.2 33 14.6 18.1 15 15 15
sum 22.7 9.5 11.5 22.4 8.1 11.5 31.9 12.5 16.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-783 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FFAGIVWQHVYEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
407
Peptide End
419
CPTAC ID
CPTAC-783
Peptide Molecular Mass
1,622.8245
Species
Homo sapiens (Human)
Assay Type
Direct MRM-MS
Matrix
Pooled patient derived xenograft breast tumor digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
TripleTOF 5600+ (SCIEX)
Internal Standard
Peptide
Peptide Standard Purity
Crude
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
2DPlus nano-LC (Eksigent)
Column Packing
ChromXP C18-CL, 3 µm, 120 Å
Column Dimensions
200 µm x 15 cm
Flow Rate
800 nL/min

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
y3 (1+) 16.3 11.1 8 16.7 13.6 7.8 23.3 17.6 11.2 15 15 15
y5 (1+) 22 9.4 7.1 27.1 11.8 8.5 34.9 15.1 11.1 15 15 15
y6 (1+) 22.9 10.8 11.8 24.8 12.9 9 33.8 16.8 14.8 15 15 15
y7 (1+) 12.8 7.2 7.6 15.2 7.3 6.5 19.9 10.3 10 15 15 15
y8 (1+) 15.5 11.6 9.5 22.3 13.1 11 27.2 17.5 14.5 15 15 15
b3 (1+) 44.5 41.7 16.3 30.3 48.7 18.4 53.8 64.1 24.6 15 15 15
sum 7.7 6.6 4.9 9.6 7.9 4.9 12.3 10.3 6.9 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3167 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FFAGIVWQHVYEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
407
Peptide End
419
CPTAC ID
CPTAC-3167
Peptide Molecular Mass
1,622.8245
Species
Homo Sapiens
Assay Type
Direct MRM-MS
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

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+) 12.6 6 7.2 11.3 8.9 11.1 16.9 10.7 13.2 15 15 15
y5 (1+) 12.1 4.4 5.8 10.8 10.1 12.7 16.2 11 14 15 15 15
y7 (1+) 4.8 4.9 6 8.1 9.7 10.7 9.4 10.9 12.3 15 15 15
sum 7.3 4.4 6 7.9 9.4 11.2 10.8 10.4 12.7 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3163 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
FYGAEIVSALDYLHSEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
252
Peptide End
268
CPTAC ID
CPTAC-3163
Peptide Molecular Mass
1,940.9520
Species
Homo Sapiens
Assay Type
Direct MRM-MS
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

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+) 4.3 4.9 5.7 6.5 4.9 6.8 7.8 6.9 8.9 15 15 15
y6 (1+) 13.5 5.4 7 9.2 4.4 8.5 16.3 7 11 15 15 15
b4 (1+) 41.2 8.2 8.5 37.5 7.8 12.5 55.7 11.3 15.1 15 15 15
sum 6.1 4.9 5.9 5.4 4.7 7.2 8.1 6.8 9.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3162 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
NDGTFIGYK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
31
Peptide End
39
CPTAC ID
CPTAC-3162
Peptide Molecular Mass
1,013.4818
Species
Homo Sapiens
Assay Type
Direct MRM-MS
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

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
y3 (1+) 13.1 11.8 8.5 13 10.3 6.5 18.5 15.7 10.7 15 15 15
y7 (1+) 11.1 9.7 8.3 11.4 9.3 7.2 15.9 13.4 11 15 15 15
y4 (1+) 13.1 12.7 7.3 13.9 11.5 6.5 19.1 17.1 9.8 15 15 15
sum 11.7 11.4 8 12 10.2 6.4 16.8 15.3 10.2 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-784 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
QEEEEMDFR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
113
Peptide End
121
CPTAC ID
CPTAC-784
Peptide Molecular Mass
1,211.4765
Species
Homo sapiens (Human)
Assay Type
Direct MRM-MS
Matrix
Pooled patient derived xenograft breast tumor digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
TripleTOF 5600+ (SCIEX)
Internal Standard
Peptide
Peptide Standard Purity
Crude
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
2DPlus nano-LC (Eksigent)
Column Packing
ChromXP C18-CL, 3 µm, 120 Å
Column Dimensions
200 µm x 15 cm
Flow Rate
800 nL/min

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+) 14.1 12.2 9.1 14.1 6.5 10.7 19.9 13.8 14 15 15 15
y5 (1+) 16.4 7.1 8.2 19 10.8 10.7 25.1 12.9 13.5 15 15 15
y6 (1+) 15.1 9.1 7.3 16.2 8.2 8.8 22.1 12.2 11.4 15 15 15
y7 (1+) 14.6 10 7.8 15.6 9.8 8.1 21.4 14 11.2 15 15 15
sum 10.1 4.1 4.5 10.2 5.2 6 14.4 6.6 7.5 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3165 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Sequence
RPHFPQFSYSASGTA
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
466
Peptide End
480
CPTAC ID
CPTAC-3165
Peptide Molecular Mass
1,651.7743
Species
Homo Sapiens
Assay Type
Direct MRM-MS
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N F
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

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
b4 (1+) 4 5.5 6 7.9 6 6.9 8.9 8.1 9.1 15 15 15
b6 (2+) 4.9 7.9 4.8 12.4 7.6 5.5 13.3 11 7.3 15 15 15
b6 (1+) 8.3 5.7 7.2 9.7 9.1 8.1 12.8 10.7 10.8 15 15 15
sum 3.2 6 5.5 8.3 6.3 6.3 8.9 8.7 8.4 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3166 Collapse assay details

Data source: Panorama

Official Gene Symbol
AKT1
Peptide Modified Sequence
TFC[+57.0]GTPEYLAPEVLEDNDYGR
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
311
Peptide - Site of Modification
3
Peptide Start
308
Peptide End
328
CPTAC ID
CPTAC-3166
Peptide Molecular Mass
2,445.0795
Species
Homo Sapiens
Assay Type
Direct MRM-MS
Matrix
Tumor Digest
Submitting Laboratory
Washington University in St. Louis
Submitting Lab PI
Reid Townsend

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
ThermoFisher, Q-Exactive
Internal Standard
25 fmol on column
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Easy-nLC 1000 Thermo Scientific
Column Packing
C18
Column Dimensions
75 µm x 50 cm
Flow Rate
300 nL/min

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 (1+) 9.3 3.3 3.7 14 7.5 6.1 16.8 8.2 7.1 15 15 15
y5 (1+) 14.3 5.1 7.5 19.6 7.2 8.3 24.3 8.8 11.2 15 15 15
b8 (1+) 22.6 5.9 6.2 25.9 9.9 8.3 34.4 11.5 10.4 15 15 15
sum 8.9 3.6 4.6 8.9 6.6 6 12.6 7.5 7.6 15 15 15


Additional Resources and Comments