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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
RB1 N/A
Sequence Length (AA) Molecular Weight (Da)
928 106159
Protein Name
Retinoblastoma-associated protein
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MPPKTPRKTA ATAAAAAAEP PAPPPPPPPE EDPEQDSGPE DLPLVRLEFE
60 70 80 90 100
ETEEPDFTAL CQKLKIPDHV RERAWLTWEK VSSVDGVLGG YIQKKKELWG
110 120 130 140 150
ICIFIAAVDL DEMSFTFTEL QKNIEISVHK FFNLLKEIDT STKVDNAMSR
160 170 180 190 200
LLKKYDVLFA LFSKLERTCE LIYLTQPSSS ISTEINSALV LKVSWITFLL
210 220 230 240 250
AKGEVLQMED DLVISFQLML CVLDYFIKLS PPMLLKEPYK TAVIPINGSP
260 270 280 290 300
RTPRRGQNRS ARIAKQLEND TRIIEVLCKE HECNIDEVKN VYFKNFIPFM
310 320 330 340 350
NSLGLVTSNG LPEVENLSKR YEEIYLKNKD LDARLFLDHD KTLQTDSIDS
360 370 380 390 400
FETQRTPRKS NLDEEVNVIP PHTPVRTVMN TIQQLMMILN SASDQPSENL
410 420 430 440 450
ISYFNNCTVN PKESILKRVK DIGYIFKEKF AKAVGQGCVE IGSQRYKLGV
460 470 480 490 500
RLYYRVMESM LKSEEERLSI QNFSKLLNDN IFHMSLLACA LEVVMATYSR
510 520 530 540 550
STSQNLDSGT DLSFPWILNV LNLKAFDFYK VIESFIKAEG NLTREMIKHL
560 570 580 590 600
ERCEHRIMES LAWLSDSPLF DLIKQSKDRE GPTDHLESAC PLNLPLQNNH
610 620 630 640 650
TAADMYLSPV RSPKKKGSTT RVNSTANAET QATSAFQTQK PLKSTSLSLF
660 670 680 690 700
YKKVYRLAYL RLNTLCERLL SEHPELEHII WTLFQHTLQN EYELMRDRHL
710 720 730 740 750
DQIMMCSMYG ICKVKNIDLK FKIIVTAYKD LPHAVQETFK RVLIKEEEYD
760 770 780 790 800
SIIVFYNSVF MQRLKTNILQ YASTRPPTLS PIPHIPRSPY KFPSSPLRIP
810 820 830 840 850
GGNIYISPLK SPYKISEGLP TPTKMTPRSR ILVSIGESFG TSEKFQKINQ
860 870 880 890 900
MVCNSDRVLK RSAEGSNPPK PLKKLRFDIE GSDEADGSKH LPGESKFQQK
910 920 928
LAEMTSTRTR MQKQKMNDSM DTSNKEEK

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-1328 Collapse assay details

Data source: Panorama

Official Gene Symbol
RB1
Peptide Modified Sequence
TAVIPINGS[+80.0]PR
Modification Type
Phospho (ST)
Protein - Site of Modification
249
Peptide - Site of Modification
9
Peptide Start
241
Peptide End
251
CPTAC ID
CPTAC-1328
Peptide Molecular Mass
1,203.6013
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
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 R
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 CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y8 (1+) 10.5 4.9 4.9 11.9 8.7 13.3 15.9 10 14.2 15 15 15
y7 (1+) 5.2 2.9 3.3 7.6 4.1 8.5 9.2 5 9.1 15 15 15
y7-98 (1+) 5.7 4.7 7.4 6.3 7.6 13.5 8.5 8.9 15.4 15 15 15
sum 4.7 2.9 3.6 5.9 4.7 9.7 7.5 5.5 10.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-942 Collapse assay details

Data source: Panorama

Official Gene Symbol
RB1
Peptide Modified Sequence
IPGGNIYIS[+80.0]PLK
Modification Type
Phospho (ST)
Protein - Site of Modification
807
Peptide - Site of Modification
9
Peptide Start
799
Peptide End
810
CPTAC ID
CPTAC-942
Peptide Molecular Mass
1,350.6949
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 CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y10 (2+) 51.1 30.6 47.9 41.2 25.3 34.9 65.6 39.7 59.3 15 24 15
y11 (2+) 50.3 15.7 16.4 40.4 12.9 17.1 64.5 20.3 23.7 15 24 15
y6 (1+) 46.2 33.1 45.2 48.9 36.1 73 67.3 49 85.9 15 24 15
sum 40.5 15.3 21 36.9 11.9 20.8 54.8 19.4 29.6 15 24 15


Additional Resources and Comments


Assay Details for CPTAC-3251 Collapse assay details

Data source: Panorama

Official Gene Symbol
RB1
Peptide Sequence
IPGGNIYISPLK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
799
Peptide End
810
CPTAC ID
CPTAC-3251
Peptide Molecular Mass
1,270.7285
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 CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y3 (1+) 14.9 12.4 10.3 18.4 14.3 12.5 23.7 18.9 16.2 15 15 15
y6 (1+) 24 12.1 5.8 22.2 13 7.3 32.7 17.8 9.3 15 15 15
y5 (1+) 26.4 18.6 16.4 36.5 18.1 18.9 45 26 25 15 15 15
sum 10 11 6 13.2 9.8 8.6 16.6 14.7 10.5 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-3288 Collapse assay details

Data source: Panorama

Official Gene Symbol
RB1
Peptide Modified Sequence
IPGGNIYIS[+80.0]PLK
Modification Type
Phospho (ST)
Protein - Site of Modification
807
Peptide - Site of Modification
9
Peptide Start
799
Peptide End
810
CPTAC ID
CPTAC-3288
Peptide Molecular Mass
1,350.6949
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 CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y5 (1+) 63.2 45.5 54.1 91.1 51.7 76.7 110.9 68.9 93.9 15 15 15
y6 (1+) 55.9 12.9 26.5 52.8 21.5 23.7 76.9 25.1 35.6 15 15 15
y10 (1+) 31.5 48.1 5.9 47 88.4 13.1 56.6 100.6 14.4 15 15 15
y10 (2+) 26.4 38.8 27.1 40 37.6 32.7 47.9 54 42.5 15 15 15
y3 (1+) 37.8 30.1 12.6 53.1 36.4 21.6 65.2 47.2 25 15 15 15
sum 19.1 12.7 9.3 33.6 18.1 12.3 38.6 22.1 15.4 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-941 Collapse assay details

Data source: Panorama

Official Gene Symbol
RB1
Peptide Modified Sequence
TAVIPINGS[+80.0]PR
Modification Type
Phospho (ST)
Protein - Site of Modification
249
Peptide - Site of Modification
9
Peptide Start
241
Peptide End
251
CPTAC ID
CPTAC-941
Peptide Molecular Mass
1,203.6013
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 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
y7 (2+) 19.7 5.9 9.9 27.5 6.8 9.9 33.8 9 14 15 24 15
y6 (1+) 18.7 9.4 9.9 32.8 12.6 13.3 37.8 15.7 16.6 15 24 15
y7 (1+) 15.6 6.5 4.8 27.1 8.5 5.8 31.3 10.7 7.5 15 24 15
sum 15.6 5.7 4.8 26.5 7.8 5.8 30.8 9.7 7.5 15 24 15


Additional Resources and Comments


Assay Details for CPTAC-898 Collapse assay details

Data source: Panorama

Official Gene Symbol
RB1
Peptide Modified Sequence
IPGGNIYIS[+80.0]PLK
Modification Type
Phospho (ST)
Protein - Site of Modification
807
Peptide - Site of Modification
9
Peptide Start
799
Peptide End
810
CPTAC ID
CPTAC-898
Peptide Molecular Mass
1,350.6949
Species
Homo sapiens (Human)
Assay Type
Direct MRM
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 at C-terminus K
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 CV
equation
n=
Fragment ion / Transition Low Med High Low Med High Low Med High Low Med High
y4 (1+) 43.6 26.7 10.8 44.5 26.2 11.4 62.3 37.4 15.7 15 15 15
b6 (1+) 36.7 29.9 18.8 43.8 28.5 19.6 57.1 41.3 27.2 15 15 15
b5 (1+) 24.3 10.6 11.2 23.8 12.8 10 34 16.6 15 15 15 15
sum 20.3 12.5 8.7 22.2 13.4 7.6 30.1 18.3 11.6 15 15 15


Additional Resources and Comments