CHEK1, CPTAC-922 - CPTAC Assay Portal (original) (raw)
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.
Protein Sequence hover to view complete sequence
| 10 | 20 | 30 | 40 | 50 |
|---|---|---|---|---|
| MAVPFVEDWD | LVQTLGEGAY | GEVQLAVNRV | TEEAVAVKIV | DMKRAVDCPE |
| 60 | 70 | 80 | 90 | 100 |
| NIKKEICINK | MLNHENVVKF | YGHRREGNIQ | YLFLEYCSGG | ELFDRIEPDI |
| 110 | 120 | 130 | 140 | 150 |
| GMPEPDAQRF | FHQLMAGVVY | LHGIGITHRD | IKPENLLLDE | RDNLKISDFG |
| 160 | 170 | 180 | 190 | 200 |
| LATVFRYNNR | ERLLNKMCGT | LPYVAPELLK | RREFHAEPVD | VWSCGIVLTA |
| 210 | 220 | 230 | 240 | 250 |
| MLAGELPWDQ | PSDSCQEYSD | WKEKKTYLNP | WKKIDSAPLA | LLHKILVENP |
| 260 | 270 | 280 | 290 | 300 |
| SARITIPDIK | KDRWYNKPLK | KGAKRPRVTS | GGVSESPSGF | SKHIQSNLDF |
| 310 | 320 | 330 | 340 | 350 |
| SPVNSASSEE | NVKYSSSQPE | PRTGLSLWDT | SPSYIDKLVQ | GISFSQPTCP |
| 360 | 370 | 380 | 390 | 400 |
| DHMLLNSQLL | GTPGSSQNPW | QRLVKRMTRF | FTKLDADKSY | QCLKETCEKL |
| 410 | 420 | 430 | 440 | 450 |
| GYQWKKSCMN | QVTISTTDRR | NNKLIFKVNL | LEMDDKILVD | FRLSKGDGLE |
| 460 | 470 | 476 | ||
| FKRHFLKIKG | KLIDIVSSQK | IWLPAT |
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-5888 Collapse assay details
Data source: Panorama
Official Gene Symbol
CHEK1
Peptide Modified Sequence
VTSGGVSES[+79.966331]PSGFSK
Modification Type
Phospho (ST)
Protein - Site of Modification
287
Peptide - Site of Modification
9
Peptide Start
278
Peptide End
292
CPTAC ID
CPTAC-5888
Peptide Molecular Mass
1,504.6447
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center
Submitting Lab PI
Amanda Paulovich
Publication
View Details (opens in a new window)
Targeted Mass Spectrometry Enables Quantification of Novel Pharmacodynamic Biomarkers of ATM Kinase Inhibition. Whiteaker JR, Wang T, Zhao L, Schoenherr RM, Kennedy JJ, Voytovich U, Ivey RG, Huang D, Lin C, Colantonio S, Caceres TW, Roberts RR, Knotts JG, Kaczmarczyk JA, Blonder J, Reading JJ, Richardson CW, Hewitt SM, Garcia-Buntley SS, Bocik W, Hiltke T, Rodriguez H, Harrington EA, Barrett JC, Lombardi B, Marco-Casanova P, Pierce AJ, Paulovich AG. Cancers (Basel). 2021 Jul 30;13(15):3843. doi: 10.3390/cancers13153843. PMID: 34359745
Assay Parameters Collapse assay parameters
Data source: Panorama
Instrument
Sciex 5500 QTRAP
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Eksigent 425
Column Packing
Reprosil
Column Dimensions
75um x 15cm
Flow Rate
300 nL/min
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 |
| y9 (1+) | 40.3 | 44.2 | 44.5 | 41.1 | 45.1 | 59 | 57.6 | 63.1 | 73.9 | 15 | 27 | 15 |
| y12 (1+) | 76.4 | 68.8 | 52.7 | 88.1 | 78.3 | 51.3 | 116.6 | 104.2 | 73.5 | 13 | 24 | 13 |
| y9-98 (1+) | 36.5 | 49.2 | 43.5 | 44.4 | 57.3 | 38.3 | 57.5 | 75.5 | 58 | 15 | 26 | 15 |
| y6 (1+) | 32.1 | 45.4 | 44.1 | 42 | 52 | 79.7 | 52.9 | 69 | 91.1 | 15 | 27 | 15 |
| sum | 8.6 | 17.9 | 15.4 | 22.3 | 15 | 19.5 | 23.9 | 23.4 | 24.8 | 15 | 27 | 15 |
Additional Resources and Comments
Assay Details for CPTAC-922 Collapse assay details
Data source: Panorama
Official Gene Symbol
CHEK1
Peptide Modified Sequence
YSSS[+80.0]QPEPR
Modification Type
Phospho (ST)
Protein - Site of Modification
317
Peptide - Site of Modification
4
Peptide Start
314
Peptide End
322
CPTAC ID
CPTAC-922
Peptide Molecular Mass
1,129.4441
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
Matrix
cell lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center
Submitting Lab PI
Amanda Paulovich
Publication
View Details (opens in a new window)
Kennedy JJ, Yan P, Zhao L, Ivey RG, Voytovich U, Moore HD, Lin C, Pogosova-Agadjanyan EL, Stirewalt DL, Reding KW, Whiteaker JR, Paulovich AG. Immobilized metal affinity chromatography coupled to multiple reaction monitoring enables reproducible quantification of phospho-signaling. Molecular and Cellular Proteomics. mcp.O115.054940
Assay Parameters Collapse assay parameters
Data source: Panorama
Instrument
QTrap 5500 (Sciex)
Internal Standard
Peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
nanoLC-2D (Eksigent)
Column Packing
ChromXP C18-CL, 3 µm, 120 Å
Column Dimensions
75 µm x 15cm
Flow Rate
300 nL/min
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 |
| y4 (1+) | 16.6 | 13.7 | 8.1 | 25.6 | 14.9 | 6.9 | 30.5 | 20.2 | 10.6 | 12 | 19 | 8 |
| y7 (1+) | 16.9 | 12.5 | 9.7 | 23.9 | 10.6 | 15.5 | 29.3 | 16.4 | 18.3 | 12 | 19 | 8 |
| b5-98 (1+) | 46.6 | 38.9 | 15.6 | 64.4 | 52 | 56 | 79.5 | 64.9 | 58.1 | 12 | 19 | 8 |
| sum | 24.7 | 15.7 | 8.1 | 32.1 | 22 | 17.4 | 40.5 | 27 | 19.2 | 12 | 19 | 8 |
Additional Resources and Comments
Assay Details for CPTAC-3223 Collapse assay details
Data source: Panorama
Official Gene Symbol
CHEK1
Peptide Sequence
YSSSQPEPR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
314
Peptide End
322
CPTAC ID
CPTAC-3223
Peptide Molecular Mass
1,049.4778
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
Matrix
cell lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center
Submitting Lab PI
Amanda Paulovich
Publication
View Details (opens in a new window)
A Multiplexed Mass Spectrometry-Based Assay for Robust Quantification of Phosphosignaling in Response to DNA Damage. Whiteaker JR, Zhao L, Saul R, Kaczmarczyk JA, Schoenherr RM, Moore HD, Jones-Weinert C, Ivey RG, Lin C, Hiltke T, Reding KW, Whiteley G, Wang P, Paulovich AG. Radiat Res. 2018 Feb 23. doi: 10.1667/RR14963.1. [Epub ahead of print]
Assay Parameters Collapse assay parameters
Data source: Panorama
Instrument
6500 QTRAP (Sciex)
Internal Standard
synthetic peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Ultra NanoLC 2D+ (Eksigent)
Column Packing
ReproSil-Pur C18-AQ 3um
Column Dimensions
100 x 0.075 mm
Flow Rate
300 uL/min
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+) | 47.2 | 26.7 | 34.4 | 51.8 | 29.2 | 32.9 | 70.1 | 39.6 | 47.6 | 15 | 15 | 15 |
| y5 (1+) | 38.7 | 22.7 | 23.5 | 43.2 | 25.7 | 23.3 | 58 | 34.3 | 33.1 | 15 | 15 | 15 |
| y7 (1+) | 29.1 | 11.2 | 15.9 | 28.6 | 20.6 | 14.6 | 40.8 | 23.4 | 21.6 | 15 | 15 | 15 |
| sum | 22.7 | 10.4 | 17.6 | 24.7 | 16.8 | 16.7 | 33.5 | 19.8 | 24.3 | 15 | 15 | 15 |
Additional Resources and Comments
Assay Details for CPTAC-3282 Collapse assay details
Data source: Panorama
Official Gene Symbol
CHEK1
Peptide Modified Sequence
YSSS[+80.0]QPEPR
Modification Type
Phospho (ST)
Protein - Site of Modification
317
Peptide - Site of Modification
4
Peptide Start
314
Peptide End
322
CPTAC ID
CPTAC-3282
Peptide Molecular Mass
1,129.4441
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
Matrix
cell lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center
Submitting Lab PI
Amanda Paulovich
Publication
View Details (opens in a new window)
A Multiplexed Mass Spectrometry-Based Assay for Robust Quantification of Phosphosignaling in Response to DNA Damage. Whiteaker JR, Zhao L, Saul R, Kaczmarczyk JA, Schoenherr RM, Moore HD, Jones-Weinert C, Ivey RG, Lin C, Hiltke T, Reding KW, Whiteley G, Wang P, Paulovich AG. Radiat Res. 2018 Feb 23. doi: 10.1667/RR14963.1. [Epub ahead of print]
Assay Parameters Collapse assay parameters
Data source: Panorama
Instrument
6500 QTRAP (Sciex)
Internal Standard
synthetic peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Ultra NanoLC 2D+ (Eksigent)
Column Packing
ReproSil-Pur C18-AQ 3um
Column Dimensions
100 x 0.075 mm
Flow Rate
300 uL/min
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+) | 36.4 | 23.8 | 14.6 | 41.7 | 31.9 | 17.9 | 55.4 | 39.8 | 23.1 | 15 | 15 | 15 |
| y6-98 (1+) | 21.6 | 17.8 | 12.1 | 39.3 | 39.9 | 35.2 | 44.8 | 43.7 | 37.2 | 15 | 15 | 15 |
| y5 (1+) | 18 | 21.9 | 33.7 | 24.3 | 37.6 | 45 | 30.2 | 43.5 | 56.2 | 15 | 15 | 15 |
| y4 (1+) | 12.8 | 6.3 | 17.6 | 23.3 | 17.8 | 20.6 | 26.6 | 18.9 | 27.1 | 15 | 15 | 15 |
| sum | 9.4 | 5.3 | 14.7 | 22.3 | 21.6 | 20.4 | 24.2 | 22.2 | 25.1 | 15 | 15 | 15 |