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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
CHEK1 CHK1
Sequence Length (AA) Molecular Weight (Da)
476 54434
Protein Name
Serine/threonine-protein kinase Chk1
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

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



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
peptide immunoaffinity
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

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+) 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
peptide immunoaffinity
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

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+) 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
peptide immunoaffinity
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

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+) 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
peptide immunoaffinity
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

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+) 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


Additional Resources and Comments