JUN, CPTAC-807 - 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
MTAKMETTFY DDALNASFLP SESGPYGYSN PKILKQSMTL NLADPVGSLK
60 70 80 90 100
PHLRAKNSDL LTSPDVGLLK LASPELERLI IQSSNGHITT TPTPTQFLCP
110 120 130 140 150
KNVTDEQEGF AEGFVRALAE LHSQNTLPSV TSAAQPVNGA GMVAPAVASV
160 170 180 190 200
AGGSGSGGFS ASLHSEPPVY ANLSNFNPGA LSSGGGAPSY GAAGLAFPAQ
210 220 230 240 250
PQQQQQPPHH LPQQMPVQHP RLQALKEEPQ TVPEMPGETP PLSPIDMESQ
260 270 280 290 300
ERIKAERKRM RNRIAASKCR KRKLERIARL EEKVKTLKAQ NSELASTANM
310 320 330 331
LREQVAQLKQ KVMNHVNSGC QLMLTQQLQT F

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-1324 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Modified Sequence

NSDLLTS[+80.0]PDVGLLK

Modification Type

Phospho (ST)

Protein - Site of Modification

63

Peptide - Site of Modification

7

Peptide Start

57

Peptide End

70

CPTAC ID

CPTAC-1324

Peptide Molecular Mass

1,550.7593

Species

Homo sapiens (Human)

Assay Type

Enrichment PRM

Enrichment Method

N/A

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 K

LC

Easy NanoLC1000

Column Packing

Reprosil C18, 3 um, 200A

Column Dimensions

0.075 x 100 mm

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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y10 (1+) | 14 | 17 | 13.3 | 19.2 | 16.1 | 19.1 | 23.8 | 23.4 | 23.3 | 15 | 15 | 15 | | y9 (1+) | 17.8 | 21.6 | 9.6 | 18.1 | 29.1 | 13 | 25.4 | 36.2 | 16.2 | 15 | 15 | 15 | | y7 (1+) | 20 | 11.1 | 8.4 | 23.9 | 12.6 | 13.8 | 31.2 | 16.8 | 16.2 | 15 | 15 | 15 | | sum | 11 | 6.4 | 6.4 | 13.6 | 11.2 | 11.7 | 17.5 | 12.9 | 13.3 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for non-CPTAC-6257 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Modified Sequence

LAS[+79.966331]PELER

Modification Type

Phospho (ST)

Protein - Site of Modification

N/A

Peptide - Site of Modification

8

Peptide Start

71

Peptide End

78

CPTAC ID

non-CPTAC-6257

Peptide Molecular Mass

993.4532

Species

Human

Assay Type

Enrichment PRM

Enrichment Method

N/A

Matrix

SKBR3 cells

Submitting Laboratory

Cell Signaling Technology

Submitting Lab PI

Yiying Zhu


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument

Orbitrap Q Exactive (Thermo)

Internal Standard

Heavy isotope-labeled peptides

Peptide Standard Purity

>95%

Peptide Standard Label Type

13C and 15N

LC

EASY-nLC II (Thermo)

Column Packing

self-packed

Column Dimensions

ID 100um, length 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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y5 (1+) | 4.2 | 6.1 | 12.3 | 4.7 | 7.9 | 11.1 | 6.3 | 10 | 16.6 | 15 | 15 | 15 | | y7-98 (1+) | 4 | 6.4 | 11.8 | 4.7 | 8.1 | 10.5 | 6.2 | 10.3 | 15.8 | 15 | 15 | 15 | | y6-98 (1+) | 5.5 | 7.3 | 11.5 | 5.4 | 7.9 | 10.1 | 7.7 | 10.8 | 15.3 | 15 | 15 | 15 | | y6-98 (2+) | 4 | 5.9 | 10.8 | 4.1 | 6.8 | 9.5 | 5.7 | 9 | 14.4 | 15 | 15 | 15 | | y7-98 (2+) | 2.9 | 7.9 | 11.6 | 3.3 | 8.8 | 10 | 4.4 | 11.8 | 15.3 | 15 | 15 | 15 | | sum | 3.6 | 6.4 | 11.4 | 3.6 | 7.5 | 10.1 | 5.1 | 9.9 | 15.2 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-927 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Modified Sequence

NSDLLTS[+80.0]PDVGLLK

Modification Type

Phospho (ST)

Protein - Site of Modification

63

Peptide - Site of Modification

7

Peptide Start

57

Peptide End

70

CPTAC ID

CPTAC-927

Peptide Molecular Mass

1,550.7593

Species

Homo sapiens (Human)

Assay Type

Enrichment PRM

Enrichment Method

N/A

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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y7 (1+) | 70.2 | 16.5 | 11.5 | 63.3 | 19.1 | 18.1 | 94.5 | 25.2 | 21.4 | 15 | 24 | 15 | | y8 (1+) | 41.2 | 22 | 15.9 | 52.4 | 24.9 | 23.2 | 66.7 | 33.2 | 28.1 | 15 | 24 | 15 | | y9 (1+) | 51.9 | 14.5 | 15.8 | 42.5 | 21.3 | 15.4 | 67.1 | 25.8 | 22.1 | 15 | 24 | 15 | | sum | 45.8 | 14 | 12.2 | 41.4 | 18.5 | 13.1 | 61.7 | 23.2 | 17.9 | 15 | 24 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-3232 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Sequence

NSDLLTSPDVGLLK

Modification Type

unmodified

Protein - Site of Modification

N/A

Peptide - Site of Modification

N/A

Peptide Start

57

Peptide End

70

CPTAC ID

CPTAC-3232

Peptide Molecular Mass

1,470.7930

Species

Homo sapiens (Human)

Assay Type

Enrichment PRM

Enrichment Method

N/A

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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y10 (1+) | 18.1 | 14.6 | 16 | 20.2 | 14.3 | 20.9 | 27.1 | 20.4 | 26.3 | 15 | 15 | 15 | | y8 (1+) | 13.8 | 16.5 | 12.6 | 19.8 | 25.3 | 20.3 | 24.1 | 30.2 | 23.9 | 15 | 15 | 15 | | y9 (1+) | 12.6 | 9.8 | 21.3 | 15.9 | 10.4 | 30.4 | 20.3 | 14.3 | 37.1 | 15 | 15 | 15 | | sum | 11.5 | 6.7 | 15.6 | 14 | 8.7 | 23.9 | 18.1 | 11 | 28.5 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-3233 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Modified Sequence

NSDLLTS[+80.0]PDVGLLK

Modification Type

Phospho (ST)

Protein - Site of Modification

63

Peptide - Site of Modification

7

Peptide Start

57

Peptide End

70

CPTAC ID

CPTAC-3233

Peptide Molecular Mass

1,550.7593

Species

Homo sapiens (Human)

Assay Type

Enrichment PRM

Enrichment Method

N/A

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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y10 (1+) | 17.5 | 10.6 | 11.9 | 32.7 | 18.6 | 16.3 | 37.1 | 21.4 | 20.2 | 15 | 15 | 15 | | y8 (1+) | 32.1 | 11.9 | 7.4 | 30.2 | 25.9 | 14.8 | 44.1 | 28.5 | 16.5 | 15 | 15 | 15 | | y9 (1+) | 23.1 | 6.7 | 13.1 | 28.1 | 14.8 | 14.9 | 36.4 | 16.2 | 19.8 | 15 | 15 | 15 | | y9-98 (1+) | 38.4 | 17 | 10.8 | 55.6 | 19.6 | 16.4 | 67.6 | 25.9 | 19.6 | 15 | 15 | 15 | | sum | 13.6 | 5.6 | 7.8 | 20.1 | 16.4 | 13 | 24.3 | 17.3 | 15.2 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-1222 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Sequence

NSDLLTSPDVGLLK

Modification Type

unmodified

Protein - Site of Modification

N/A

Peptide - Site of Modification

N/A

Peptide Start

57

Peptide End

70

CPTAC ID

CPTAC-1222

Peptide Molecular Mass

1,470.7930

Species

Homo sapiens (Human)

Assay Type

Direct PRM

Matrix

Ovarian cancer tumor tissue lysate

Submitting Laboratory

Pacific Northwest National Laboratory

Submitting Lab PI

Tao Liu


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument

TSQ Vantage

Internal Standard

peptide

Peptide Standard Purity

>95%

Peptide Standard Label Type

13C and 15N at C-terminus K

LC

Waters nanoACQUITY (Part Number 176016000)

Column Packing

Waters BEH C18, 1.7 um 130 Å (Part Number 186007485)

Column Dimensions

100 um x 100 mm

Flow Rate

0.5 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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y4 (1+) | 32.6 | 6.6 | 7.4 | 34.4 | 11.6 | 13 | 47.4 | 13.3 | 15 | 15 | 15 | 15 | | y7 (1+) | 9 | 5.7 | 5.1 | 19.5 | 13.3 | 11.5 | 21.5 | 14.5 | 12.6 | 15 | 15 | 15 | | y9 (1+) | 7.7 | 4.5 | 5.4 | 12.2 | 10 | 11.9 | 14.4 | 11 | 13.1 | 15 | 15 | 15 | | sum | 8 | 4.9 | 5.2 | 13.7 | 11.5 | 11.7 | 15.9 | 12.5 | 12.8 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-807 Collapse assay details

Data source: Panorama

Official Gene Symbol

JUN

Peptide Modified Sequence

LAS[+80.0]PELER

Modification Type

Phospho (ST)

Protein - Site of Modification

73

Peptide - Site of Modification

3

Peptide Start

71

Peptide End

78

CPTAC ID

CPTAC-807

Peptide Molecular Mass

993.4532

Species

Homo sapiens (Human)

Assay Type

Direct PRM

Matrix

Pooled patient derived xenograft breast tumor digest

Submitting Laboratory

Washington University in St. Louis

Submitting Lab PI

Reid Townsend


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument

TripleTOF 5600+ (SCIEX)

Internal Standard

Peptide

Peptide Standard Purity

Crude

Peptide Standard Label Type

13C and 15N

LC

2DPlus nano-LC (Eksigent)

Column Packing

ChromXP C18-CL, 3 µm, 120 Å

Column Dimensions

200 µm x 15 cm

Flow Rate

800 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 CVequation | n= | | | | | | | | | | | ---------------------------------------- | -------------------------------------- | --------------------------------------------------------------------------------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | ------- | ------- | -------- | | Fragment ion / Transition | Low | Med | High | Low | Med | High | Low | Med | High | Low | Med | High | | y3 (1+) | 12.9 | 3.5 | 3.3 | 13.1 | 4.3 | 3.9 | 18.4 | 5.5 | 5.1 | 15 | 15 | 15 | | y4 (1+) | 7.7 | 4.2 | 4.4 | 10.8 | 4 | 3.9 | 13.3 | 5.8 | 5.9 | 15 | 15 | 15 | | y5 (1+) | 8 | 2.3 | 3.1 | 9.5 | 2.9 | 3.7 | 12.4 | 3.7 | 4.8 | 15 | 15 | 15 | | b3 (1+) | 23.9 | 11.4 | 8.9 | 20 | 11.3 | 7.9 | 31.2 | 16.1 | 11.9 | 15 | 15 | 15 | | sum | 6.6 | 2 | 2.2 | 6.7 | 2.3 | 2.8 | 9.4 | 3 | 3.6 | 15 | 15 | 15 |



Additional Resources and Comments