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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
RAD18 RNF73
Sequence Length (AA) Molecular Weight (Da)
495 56223
Protein Name
E3 ubiquitin-protein ligase RAD18
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MDSLAESRWP PGLAVMKTID DLLRCGICFE YFNIAMIIPQ CSHNYCSLCI
60 70 80 90 100
RKFLSYKTQC PTCCVTVTEP DLKNNRILDE LVKSLNFARN HLLQFALESP
110 120 130 140 150
AKSPASSSSK NLAVKVYTPV ASRQSLKQGS RLMDNFLIRE MSGSTSELLI
160 170 180 190 200
KENKSKFSPQ KEASPAAKTK ETRSVEEIAP DPSEAKRPEP PSTSTLKQVT
210 220 230 240 250
KVDCPVCGVN IPESHINKHL DSCLSREEKK ESLRSSVHKR KPLPKTVYNL
260 270 280 290 300
LSDRDLKKKL KEHGLSIQGN KQQLIKRHQE FVHMYNAQCD ALHPKSAAEI
310 320 330 340 350
VREIENIEKT RMRLEASKLN ESVMVFTKDQ TEKEIDEIHS KYRKKHKSEF
360 370 380 390 400
QLLVDQARKG YKKIAGMSQK TVTITKEDES TEKLSSVCMG QEDNMTSVTN
410 420 430 440 450
HFSQSKLDSP EELEPDREED SSSCIDIQEV LSSSESDSCN SSSSDIIRDL
460 470 480 490 495
LEEEEAWEAS HKNDLQDTEI SPRQNRRTRA AESAEIEPRN KRNRN

Data source: UniProt


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

Uniprot Database Entry PhosphoSitePlus ®

Click a point on a node
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-940 Collapse assay details

Data source: Panorama

Official Gene Symbol
RAD18
Peptide Modified Sequence
NDLQDTEIS[+80.0]PR
Modification Type
Phospho (ST)
Protein - Site of Modification
471
Peptide - Site of Modification
9
Peptide Start
463
Peptide End
473
CPTAC ID
CPTAC-940
Peptide Molecular Mass
1,366.5766
Species
Homo sapiens (Human)
Assay Type
Enrichment ImmunoMRM
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
y9 (2+) 21.1 11.7 18.4 22 14.7 21.6 30.5 18.8 28.4 15 24 15
y6 (1+) 17.6 17 13.8 26.9 21.5 15.7 32.1 27.4 20.9 15 24 15
y7 (1+) 14.9 8.7 11 26.7 12.5 11.4 30.6 15.2 15.8 15 24 15
sum 11.5 9.2 11 23.8 13.9 12.5 26.4 16.7 16.7 15 24 15


Additional Resources and Comments


Assay Details for CPTAC-3247 Collapse assay details

Data source: Panorama

Official Gene Symbol
RAD18
Peptide Modified Sequence
NDLQDTEIS[+80.0]PR
Modification Type
Phospho (ST)
Protein - Site of Modification
471
Peptide - Site of Modification
9
Peptide Start
463
Peptide End
473
CPTAC ID
CPTAC-3247
Peptide Molecular Mass
1,366.5766
Species
Homo sapiens (Human)
Assay Type
Enrichment ImmunoMRM
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
y8-98 (1+) 28 7.9 11.1 32.5 11.4 12.3 42.9 13.9 16.6 15 15 15
y4-98 (1+) 37.3 13.6 13.7 42.4 19.6 12.4 56.5 23.9 18.5 15 15 15
y4 (1+) 37.6 9.9 7 49.6 13 8.9 62.2 16.3 11.3 15 15 15
y7-98 (1+) 16.5 8.3 8.5 33.1 20 9 37 21.7 12.4 15 15 15
y7 (1+) 18 6.9 6.7 30.9 11.4 7.2 35.8 13.3 9.8 15 15 15
sum 12.9 5.5 5.7 30.4 11.8 6.3 33 13 8.5 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-939 Collapse assay details

Data source: Panorama

Official Gene Symbol
RAD18
Peptide Modified Sequence
NHLLQFALES[+80.0]PAK
Modification Type
Phospho (ST)
Protein - Site of Modification
99
Peptide - Site of Modification
10
Peptide Start
90
Peptide End
102
CPTAC ID
CPTAC-939
Peptide Molecular Mass
1,546.7545
Species
Homo sapiens (Human)
Assay Type
Enrichment ImmunoMRM
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 K
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
y5 (1+) 35.5 28.1 58.3 37.8 27.2 54.7 51.9 39.1 79.9 12 20 15
y6 (1+) 51 21.9 21.9 29.3 23.2 37.1 58.8 31.9 43.1 12 20 15
b7 (2+) 43.3 17.3 49.1 37.8 31.2 65.9 57.5 35.7 82.2 12 20 15
sum 42.6 13.5 22.6 22.4 15 29.8 48.1 20.2 37.4 12 20 15


Additional Resources and Comments


Assay Details for CPTAC-3246 Collapse assay details

Data source: Panorama

Official Gene Symbol
RAD18
Peptide Sequence
NHLLQFALESPAK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
90
Peptide End
102
CPTAC ID
CPTAC-3246
Peptide Molecular Mass
1,466.7882
Species
Homo sapiens (Human)
Assay Type
Enrichment ImmunoMRM
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 K
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
y10 (1+) 9.5 15.9 8.9 10.6 36.1 7.6 14.2 39.4 11.7 15 15 15
y3 (1+) 30.4 17.2 16 34.7 29 20.4 46.1 33.7 25.9 15 15 15
y9 (1+) 21.8 12 10.4 28.5 13.4 14.2 35.9 18 17.6 15 15 15
y5 (1+) 30.6 21.3 15.3 29 24.2 20.8 42.2 32.2 25.8 15 15 15
sum 10.1 11 6.3 13.1 23.8 6.8 16.5 26.2 9.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3285 Collapse assay details

Data source: Panorama

Official Gene Symbol
RAD18
Peptide Modified Sequence
NHLLQFALES[+80.0]PAK
Modification Type
Phospho (ST)
Protein - Site of Modification
99
Peptide - Site of Modification
10
Peptide Start
90
Peptide End
102
CPTAC ID
CPTAC-3285
Peptide Molecular Mass
1,546.7545
Species
Homo sapiens (Human)
Assay Type
Enrichment ImmunoMRM
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 K
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
y3 (1+) 24.6 9.9 15.3 43.3 11.7 24.9 49.8 15.3 29.2 15 15 15
y4 (1+) 37.4 19.3 15.4 41.5 21.3 19.5 55.9 28.7 24.8 15 15 15
y5 (1+) 41.7 29.3 20.4 63.7 19.1 20.2 76.1 35 28.7 15 15 15
y6 (1+) 29.9 14.9 10.8 58.2 54.5 42.2 65.4 56.5 43.6 15 15 15
sum 23.5 9.5 9.4 37.8 21.7 23.1 44.5 23.7 24.9 15 15 15


Additional Resources and Comments