FOXO1, CPTAC-1323 - CPTAC Assay Portal (original) (raw)
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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.
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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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
Repeatability
Data source: Panorama
| | Average intra-assay CV(within day CV) | Average inter-assay CV(between day CV) | Total CV
| 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
Repeatability
Data source: Panorama
| | Average intra-assay CV(within day CV) | Average inter-assay CV(between day CV) | Total CV | 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
Repeatability
Data source: Panorama
| | Average intra-assay CV(within day CV) | Average inter-assay CV(between day CV) | Total CV | 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 |