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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
FOXO1 FKHR, FOXO1A
Sequence Length (AA) Molecular Weight (Da)
655 69662
Protein Name
Forkhead box protein O1
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MAEAPQVVEI DPDFEPLPRP RSCTWPLPRP EFSQSNSATS SPAPSGSAAA
60 70 80 90 100
NPDAAAGLPS ASAAAVSADF MSNLSLLEES EDFPQAPGSV AAAVAAAAAA
110 120 130 140 150
AATGGLCGDF QGPEAGCLHP APPQPPPPGP LSQHPPVPPA AAGPLAGQPR
160 170 180 190 200
KSSSSRRNAW GNLSYADLIT KAIESSAEKR LTLSQIYEWM VKSVPYFKDK
210 220 230 240 250
GDSNSSAGWK NSIRHNLSLH SKFIRVQNEG TGKSSWWMLN PEGGKSGKSP
260 270 280 290 300
RRRAASMDNN SKFAKSRSRA AKKKASLQSG QEGAGDSPGS QFSKWPASPG
310 320 330 340 350
SHSNDDFDNW STFRPRTSSN ASTISGRLSP IMTEQDDLGE GDVHSMVYPP
360 370 380 390 400
SAAKMASTLP SLSEISNPEN MENLLDNLNL LSSPTSLTVS TQSSPGTMMQ
410 420 430 440 450
QTPCYSFAPP NTSLNSPSPN YQKYTYGQSS MSPLPQMPIQ TLQDNKSSYG
460 470 480 490 500
GMSQYNCAPG LLKELLTSDS PPHNDIMTPV DPGVAQPNSR VLGQNVMMGP
510 520 530 540 550
NSVMSTYGSQ ASHNKMMNPS SHTHPGHAQQ TSAVNGRPLP HTVSTMPHTS
560 570 580 590 600
GMNRLTQVKT PVQVPLPHPM QMSALGGYSS VSSCNGYGRM GLLHQEKLPS
610 620 630 640 650
DLDGMFIERL DCDMESIIRN DLMDGDTLDF NFDNVLPNQS FPHSVKTTTH
655
SWVSG

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
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Phosphorylation Acetylation Ubiquitylation Other

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Assay Details for CPTAC-1323 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO1
Peptide Modified Sequence
TSS[+80.0]NASTISGR
Modification Type
Phospho (ST)
Protein - Site of Modification
319
Peptide - Site of Modification
3
Peptide Start
317
Peptide End
327
CPTAC ID
CPTAC-1323
Peptide Molecular Mass
1,159.4870
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)

Abelin, et al. Mol Cell Proteomics. 2016 May;15(5):1622-41. doi: 10.1074/mcp.M116.058354. Epub 2016 Feb 24. PMID:26912667


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
medium stable isotope peptide (delta 6 amu)
Peptide Standard Purity
Crude (~60%)
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 3 um, 200A
Column Dimensions
0.075 x 100 mm
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


Chromatograms

Data source: Panorama


Response Curves

Data source: Panorama

Retrieving Data

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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+) 6.2 3.1 3.6 3.6 3.9 10.5 7.2 5 11.1 15 15 15
y7 (1+) 5.7 4.3 3.2 7.3 5 8.8 9.3 6.6 9.4 15 15 15
y6 (1+) 6.5 3.4 2.9 6.5 3.6 7.6 9.2 5 8.1 15 15 15
sum 5.2 3.1 2.6 4.6 3.5 8.3 6.9 4.7 8.7 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-6045 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO1
Peptide Sequence
ASLQSGQEGAGDSPGSQFSK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
275
Peptide End
294
CPTAC ID
CPTAC-6045
Peptide Molecular Mass
1,936.8763
Species
Human
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
FFPE lung tumor tissue lysate pool
Submitting Laboratory
Fred Hutchinson Cancer Research Center
Submitting Lab PI
Amanda Paulovich

Publication

View Details (opens in a new window)

Whiteaker JR, Zhao L, Schoenherr RM, Huang D, Lundeen RA, Voytovich U, Kennedy JJ, Ivey RG, Lin C, Murillo OD, Lorentzen TD, Colantonio S, Caceres TW, Roberts RR, Knotts JG, Reading JJ, Perry CD, Richardson CW, Garcia-Buntley SS, Bocik W, Hewitt SM, Chowdhury S, Vandermeer J, Smith SD, Gopal AK, Ramchurren N, Fling SP, Wang P, Paulovich AG. A multiplexed assay for quantifying immunomodulatory proteins supports correlative studies in immunotherapy clinical trials. Front Oncol. 2023 May 2;13:1168710. doi: 10.3389/fonc.2023.1168710. PMID: 37205196; PMCID: PMC10185886.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
QTRAP 5500
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Eksigent nanoLC 425
Column Packing
3 µm ChromXP C18EP, 120 Å
Column Dimensions
150 x 0.075 mm
Flow Rate
0.3 µL/min

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


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+) 24.4 15.6 17.4 26.9 19.5 13.6 36.3 25 22.1 15 15 15
y6 (1+) 19.9 22.3 19.9 23.2 25.4 17.7 30.6 33.8 26.6 15 15 15
y7 (1+) 20 12.5 14.3 27.1 17.9 14.5 33.7 21.8 20.4 15 15 15
y8 (1+) 25.3 19.8 14.1 26.9 24.7 13.5 36.9 31.7 19.5 15 15 15
y16 (2+) 32 21.5 20.6 31.6 27 29.2 45 34.5 35.7 15 15 15
y7 (2+) 23.4 14.5 16.9 26.7 17.8 14.5 35.5 23 22.3 15 15 15
y8 (2+) 15.2 13 17.5 14 28.4 17.9 20.7 31.2 25 15 15 15
sum 16.6 11.1 14.8 21 16.2 13.6 26.8 19.6 20.1 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5703 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO1
Peptide Modified Sequence
AAS[+79.966331]MDNNSK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
254
Peptide End
262
CPTAC ID
non-CPTAC-5703
Peptide Molecular Mass
1,016.3634
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-Biologist curated set of targets across a number of important cancer pathways


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+) 5.5 6.6 7.3 14.1 5.9 8.7 15.1 8.9 11.4 15 13 14
y7-98 (2+) 15.7 7.5 7 10.8 6.5 5.7 19.1 9.9 9 15 13 14
y8-98 (2+) 15.3 6.6 9.1 13.8 6.8 8.4 20.6 9.5 12.4 15 13 14
y5 (1+) 10.6 5.5 13.3 11.5 5.4 16.8 15.6 7.7 21.4 15 13 14
y7-98 (1+) 8.6 6.7 6.2 9 6.3 7 12.4 9.2 9.4 15 13 14
sum 10.2 5.5 9.1 9.4 4 8.9 13.9 6.8 12.7 15 13 14


Additional Resources and Comments