RRM2, CPTAC-725 - 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

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-725 Collapse assay details

Data source: Panorama

Official Gene Symbol

RRM2

Peptide Sequence

DIQHWESLKPEER

Modification Type

unmodified

Protein - Site of Modification

N/A

Peptide - Site of Modification

N/A

Peptide Start

112

Peptide End

124

CPTAC ID

CPTAC-725

Peptide Molecular Mass

1,665.8111

Species

Homo sapiens (Human)

Assay Type

Enrichment ImmunoMRM

Enrichment Method

peptide immunoaffinity

Matrix

plasma

Submitting Laboratory

Fred Hutchinson Cancer Research Center

Submitting Lab PI

Amanda Paulovich

Publication

View Details (opens in a new window)

Anti-peptide monoclonal antibodies generated for immuno-multiple reaction monitoring-mass spectrometry assays have a high probability of supporting Western blot and ELISA. Schoenherr RM, Saul RG, Whiteaker JR, Yan P, Whiteley GR, Paulovich AG. Mol Cell Proteomics. 2015 Feb;14(2):382-98. doi: 10.1074/mcp.O114.043133. Epub 2014 Dec 15. PMID: 25512614

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument

5500 Qtrap

Internal Standard

synthetic peptide

Peptide Standard Purity

>95%

Peptide Standard Label Type

13C and 15N at C-terminus R

LC

Eksigent NanoLC 2D

Column Packing

ReproSil-Pur C18-AQ 3 um

Column Dimensions

100 mm x 75 um ID

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 | | b11 (2+) | 14 | 4.5 | 8.1 | 10.5 | 5.7 | 9 | 17.5 | 7.3 | 12.1 | 15 | 15 | 15 | | y9 (2+) | 16.1 | 9 | 9.7 | 17 | 8.9 | 10.7 | 23.4 | 12.7 | 14.4 | 15 | 15 | 15 | | y11 (2+) | 9.5 | 5.6 | 4.4 | 8.1 | 8 | 5.9 | 12.5 | 9.8 | 7.4 | 15 | 15 | 15 | | y2 (1+) | 15 | 7.6 | 5.5 | 14.4 | 7.8 | 7.9 | 20.8 | 10.9 | 9.6 | 15 | 15 | 15 | | sum | 5.7 | 3.7 | 3.3 | 8.1 | 4.7 | 5.1 | 9.9 | 6 | 6.1 | 15 | 15 | 15 |

Additional Resources and Comments

Assay Details for CPTAC-3255 Collapse assay details

Data source: Panorama

Official Gene Symbol

RRM2

Peptide Sequence

IEQEFLTEALPVK

Modification Type

unmodified

Protein - Site of Modification

N/A

Peptide - Site of Modification

N/A

Peptide Start

299

Peptide End

311

CPTAC ID

CPTAC-3255

Peptide Molecular Mass

1,515.8185

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

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+) | 14.6 | 9.9 | 10.1 | 20.2 | 10.8 | 18.4 | 24.9 | 14.7 | 21 | 15 | 15 | 15 | | y9 (1+) | 11.2 | 11.3 | 13.6 | 15 | 13.2 | 11.8 | 18.7 | 17.4 | 18 | 15 | 15 | 15 | | y4 (1+) | 12.5 | 10.3 | 10.4 | 13 | 10.7 | 12.2 | 18 | 14.9 | 16 | 15 | 15 | 15 | | b3 (1+) | 20.2 | 9 | 12.7 | 19.6 | 8.7 | 16.1 | 28.1 | 12.5 | 20.5 | 15 | 15 | 15 | | sum | 7.7 | 7.3 | 7.5 | 8.1 | 7.1 | 10.2 | 11.2 | 10.2 | 12.7 | 15 | 15 | 15 |

Additional Resources and Comments