NBN, CPTAC-934 - 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
MWKLLPAAGP AGGEPYRLLT GVEYVVGRKN CAILIENDQS ISRNHAVLTA
60 70 80 90 100
NFSVTNLSQT DEIPVLTLKD NSKYGTFVNE EKMQNGFSRT LKSGDGITFG
110 120 130 140 150
VFGSKFRIEY EPLVACSSCL DVSGKTALNQ AILQLGGFTV NNWTEECTHL
160 170 180 190 200
VMVSVKVTIK TICALICGRP IVKPEYFTEF LKAVESKKQP PQIESFYPPL
210 220 230 240 250
DEPSIGSKNV DLSGRQERKQ IFKGKTFIFL NAKQHKKLSS AVVFGGGEAR
260 270 280 290 300
LITEENEEEH NFFLAPGTCV VDTGITNSQT LIPDCQKKWI QSIMDMLQRQ
310 320 330 340 350
GLRPIPEAEI GLAVIFMTTK NYCDPQGHPS TGLKTTTPGP SLSQGVSVDE
360 370 380 390 400
KLMPSAPVNT TTYVADTESE QADTWDLSER PKEIKVSKME QKFRMLSQDA
410 420 430 440 450
PTVKESCKTS SNNNSMVSNT LAKMRIPNYQ LSPTKLPSIN KSKDRASQQQ
460 470 480 490 500
QTNSIRNYFQ PSTKKRERDE ENQEMSSCKS ARIETSCSLL EQTQPATPSL
510 520 530 540 550
WKNKEQHLSE NEPVDTNSDN NLFTDTDLKS IVKNSASKSH AAEKLRSNKK
560 570 580 590 600
REMDDVAIED EVLEQLFKDT KPELEIDVKV QKQEEDVNVR KRPRMDIETN
610 620 630 640 650
DTFSDEAVPE SSKISQENEI GKKRELKEDS LWSAKEISNN DKLQDDSEML
660 670 680 690 700
PKKLLLTEFR SLVIKNSTSR NPSGINDDYG QLKNFKKFKK VTYPGAGKLP
710 720 730 740 750
HIIGGSDLIA HHARKNTELE EWLRQEMEVQ NQHAKEESLA DDLFRYNPYL
754
KRRR

Data source: UniProt


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

Uniprot Database Entry PhosphoSitePlus ®

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All other points link out to UniProt


Phosphorylation Acetylation Ubiquitylation Other

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Assay Details for CPTAC-935 Collapse assay details

Data source: Panorama

Official Gene Symbol

NBN

Peptide Modified Sequence

IPNYQLS[+80.0]PTK

Modification Type

Phospho (ST)

Protein - Site of Modification

432

Peptide - Site of Modification

7

Peptide Start

426

Peptide End

435

CPTAC ID

CPTAC-935

Peptide Molecular Mass

1,239.5901

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 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 | | b7-98 (1+) | 52.6 | 14.9 | 37.6 | 52.5 | 13.6 | 31.9 | 74.3 | 20.2 | 49.3 | 15 | 24 | 15 | | y8 (2+) | 14.4 | 12 | 16.9 | 38.1 | 11.6 | 21.2 | 40.7 | 16.7 | 27.1 | 15 | 24 | 15 | | y9 (2+) | 22.8 | 7.3 | 7.8 | 29.3 | 11 | 8.9 | 37.1 | 13.2 | 11.8 | 15 | 24 | 15 | | sum | 20 | 7.2 | 8.1 | 27.9 | 10 | 10.4 | 34.3 | 12.3 | 13.2 | 15 | 24 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-3238 Collapse assay details

Data source: Panorama

Official Gene Symbol

NBN

Peptide Sequence

IPNYQLSPTK

Modification Type

unmodified

Protein - Site of Modification

N/A

Peptide - Site of Modification

N/A

Peptide Start

426

Peptide End

435

CPTAC ID

CPTAC-3238

Peptide Molecular Mass

1,159.6237

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 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 | | y7 (1+) | 19.9 | 8.5 | 7.4 | 20.3 | 8.3 | 7.6 | 28.4 | 11.9 | 10.6 | 15 | 15 | 15 | | y8 (1+) | 11.9 | 9.6 | 4.7 | 23.3 | 15.8 | 10.7 | 26.2 | 18.5 | 11.7 | 15 | 15 | 15 | | y4 (1+) | 28.5 | 10.5 | 5 | 35.1 | 14.8 | 10.2 | 45.2 | 18.1 | 11.4 | 15 | 15 | 15 | | sum | 13.6 | 7.6 | 4 | 17.8 | 12.1 | 8.8 | 22.4 | 14.3 | 9.7 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-3239 Collapse assay details

Data source: Panorama

Official Gene Symbol

NBN

Peptide Modified Sequence

IPNYQLS[+80.0]PTK

Modification Type

Phospho (ST)

Protein - Site of Modification

432

Peptide - Site of Modification

7

Peptide Start

426

Peptide End

435

CPTAC ID

CPTAC-3239

Peptide Molecular Mass

1,239.5901

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 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 | | y8 (2+) | 19 | 24.5 | 13 | 38 | 27.3 | 20.9 | 42.5 | 36.7 | 24.6 | 15 | 15 | 15 | | y8 (1+) | 18.2 | 13 | 9.8 | 42.5 | 18 | 12.1 | 46.2 | 22.2 | 15.6 | 15 | 15 | 15 | | y9-98 (2+) | 17.6 | 11.1 | 8.2 | 47.2 | 14.5 | 12.7 | 50.4 | 18.3 | 15.1 | 15 | 15 | 15 | | y8-98 (1+) | 32 | 13.3 | 11.1 | 47.2 | 13.7 | 20 | 57 | 19.1 | 22.9 | 15 | 15 | 15 | | sum | 12.6 | 11.5 | 6.8 | 42.1 | 15.3 | 12.5 | 43.9 | 19.1 | 14.2 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-934 Collapse assay details

Data source: Panorama

Official Gene Symbol

NBN

Peptide Modified Sequence

TTTPGPSLS[+80.0]QGVSVDEK

Modification Type

Phospho (ST)

Protein - Site of Modification

343

Peptide - Site of Modification

9

Peptide Start

335

Peptide End

351

CPTAC ID

CPTAC-934

Peptide Molecular Mass

1,781.8084

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 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 | | y13 (2+) | 28.6 | 8 | 10.1 | 23.1 | 11.3 | 14.8 | 36.8 | 13.8 | 17.9 | 15 | 24 | 15 | | y14 (2+) | 33.1 | 8.7 | 9.6 | 24.2 | 10.4 | 12.2 | 41 | 13.6 | 15.5 | 15 | 24 | 15 | | y15 (2+) | 48.5 | 11.4 | 8.5 | 40.3 | 11.6 | 14.1 | 63.1 | 16.3 | 16.5 | 15 | 24 | 15 | | sum | 32.1 | 8.2 | 8.3 | 20.9 | 9.6 | 12.1 | 38.3 | 12.6 | 14.7 | 15 | 24 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-3236 Collapse assay details

Data source: Panorama

Official Gene Symbol

NBN

Peptide Sequence

TTTPGPSLSQGVSVDEK

Modification Type

unmodified

Protein - Site of Modification

N/A

Peptide - Site of Modification

N/A

Peptide Start

335

Peptide End

351

CPTAC ID

CPTAC-3236

Peptide Molecular Mass

1,701.8421

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 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 | | y9 (1+) | 86.1 | 26.9 | 20.6 | 90.6 | 28.8 | 20 | 125 | 39.4 | 28.7 | 15 | 15 | 15 | | y12 (2+) | 72.5 | 49.6 | 54.3 | 64.7 | 53.6 | 76.5 | 97.2 | 73 | 93.8 | 15 | 15 | 15 | | y14 (2+) | 25.5 | 14.5 | 17.3 | 25.5 | 14.2 | 16.7 | 36.1 | 20.3 | 24 | 15 | 15 | 15 | | sum | 23.1 | 12.1 | 16.7 | 22.9 | 12.9 | 16.3 | 32.5 | 17.7 | 23.3 | 15 | 15 | 15 |



Additional Resources and Comments


Assay Details for CPTAC-3237 Collapse assay details

Data source: Panorama

Official Gene Symbol

NBN

Peptide Modified Sequence

TTTPGPSLS[+80.0]QGVSVDEK

Modification Type

Phospho (ST)

Protein - Site of Modification

343

Peptide - Site of Modification

9

Peptide Start

335

Peptide End

351

CPTAC ID

CPTAC-3237

Peptide Molecular Mass

1,781.8084

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 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 | | y13 (2+) | 28.1 | 11.5 | 20 | 39.3 | 10.8 | 16.5 | 48.3 | 15.8 | 25.9 | 15 | 15 | 15 | | y14 (2+) | 22 | 8.5 | 7.8 | 27.2 | 10.1 | 7 | 35 | 13.2 | 10.5 | 15 | 15 | 15 | | y15 (2+) | 33.1 | 10.6 | 7.9 | 38.9 | 14.7 | 13.1 | 51.1 | 18.1 | 15.3 | 15 | 15 | 15 | | y5 (1+) | 44.8 | 8.8 | 13.9 | 47 | 8.6 | 13.5 | 64.9 | 12.3 | 19.4 | 15 | 15 | 15 | | sum | 12.5 | 6.6 | 7.1 | 24.3 | 8.3 | 5.9 | 27.3 | 10.6 | 9.2 | 15 | 15 | 15 |



Additional Resources and Comments