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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
HSPB1 HSP27, HSP28
Sequence Length (AA) Molecular Weight (Da)
205 22783
Protein Name
Heat shock protein beta-1
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MTERRVPFSL LRGPSWDPFR DWYPHSRLFD QAFGLPRLPE EWSQWLGGSS
60 70 80 90 100
WPGYVRPLPP AAIESPAVAA PAYSRALSRQ LSSGVSEIRH TADRWRVSLD
110 120 130 140 150
VNHFAPDELT VKTKDGVVEI TGKHEERQDE HGYISRCFTR KYTLPPGVDP
160 170 180 190 200
TQVSSSLSPE GTLTVEAPMP KLATQSNEIT IPVTFESRAQ LGGPEAAKSD
205
ETAAK

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-1427 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
AQLGGPEAAK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
189
Peptide End
198
CPTAC ID
CPTAC-1427
Peptide Molecular Mass
940.4978
Species
Homo sapiens (Human)
Assay Type
Direct ImmunoMRM
Matrix
depleted digested plasma
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Large-Scale Interlaboratory Study to Develop, Analytically Validate and Apply Highly Multiplexed, Quantitative Peptide Assays to Measure Cancer-Relevant Proteins in Plasma. Abbatiello SE, Schilling B, Mani DR, Zimmerman LJ, Hall SC, MacLean B, Albertolle M, Allen S, Burgess M, Cusack MP, Gosh M, Hedrick V, Held JM, Inerowicz HD, Jackson A, Keshishian H, Kinsinger CR, Lyssand J, Makowski L, Mesri M, Rodriguez H, Rudnick P, Sadowski P, Sedransk N, Shaddox K, Skates SJ, Kuhn E, Smith D, Whiteaker JR, Whitwell C, Zhang S, Borchers CH, Fisher SJ, Gibson BW, Liebler DC, MacCoss MJ, Neubert TA, Paulovich AG, Regnier FE, Tempst P, Carr SA. Mol Cell Proteomics. 2015 Sep;14(9):2357-74. doi: 10.1074/mcp.M114.047050. Epub 2015 Feb 18. PMID: 25693799


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
heavy stable isotope peptide (delta 8-10)
Peptide Standard Purity
>95%
Peptide Standard Label Type
15N
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 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+) 3.2 2.2 2.3 4.4 6.2 4 5.4 6.6 4.6 15 15 15
y7 (1+) 2.5 2.5 1.8 6.4 6.5 3.6 6.9 7 4 15 15 15
y6 (1+) 6 2.5 2 11.5 5.3 3.7 13 5.9 4.2 15 15 15
sum 3 2.1 1.8 5.3 6.2 3.7 6.1 6.5 4.1 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1429 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
DGVVEITGK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
115
Peptide End
123
CPTAC ID
CPTAC-1429
Peptide Molecular Mass
916.4866
Species
Homo sapiens (Human)
Assay Type
Direct ImmunoMRM
Matrix
depleted digested plasma
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Large-Scale Interlaboratory Study to Develop, Analytically Validate and Apply Highly Multiplexed, Quantitative Peptide Assays to Measure Cancer-Relevant Proteins in Plasma. Abbatiello SE, Schilling B, Mani DR, Zimmerman LJ, Hall SC, MacLean B, Albertolle M, Allen S, Burgess M, Cusack MP, Gosh M, Hedrick V, Held JM, Inerowicz HD, Jackson A, Keshishian H, Kinsinger CR, Lyssand J, Makowski L, Mesri M, Rodriguez H, Rudnick P, Sadowski P, Sedransk N, Shaddox K, Skates SJ, Kuhn E, Smith D, Whiteaker JR, Whitwell C, Zhang S, Borchers CH, Fisher SJ, Gibson BW, Liebler DC, MacCoss MJ, Neubert TA, Paulovich AG, Regnier FE, Tempst P, Carr SA. Mol Cell Proteomics. 2015 Sep;14(9):2357-74. doi: 10.1074/mcp.M114.047050. Epub 2015 Feb 18. PMID: 25693799


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
heavy stable isotope peptide (delta 8-10)
Peptide Standard Purity
>95%
Peptide Standard Label Type
15N
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 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 (1+) 7 3.5 2.2 8.9 5.3 3.5 11.3 6.4 4.1 15 15 15
y6 (1+) 1.9 1.1 1 3.6 5.3 3 4.1 5.4 3.2 15 15 15
y5 (1+) 2.6 1.9 1.7 3.5 5.3 3.4 4.4 5.6 3.8 15 15 15
sum 1.2 1.2 0.9 3.1 5.2 3 3.3 5.3 3.1 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1351 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Modified Sequence
GPS[+80.0]WDPFRDWYPHSR
Modification Type
Phospho (ST)
Protein - Site of Modification
15
Peptide - Site of Modification
3
Peptide Start
13
Peptide End
27
CPTAC ID
CPTAC-1351
Peptide Molecular Mass
1,981.8261
Species
Homo sapiens (Human)
Assay Type
Enrichment ImmunoMRM
Enrichment Method
peptide immunoaffinity
Matrix
digested cell lysate
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Mol Cell Proteomics. 2016 May;15(5):1622-41. doi: 10.1074/mcp.M116.058354. Epub 2016 Feb 24. Reduced-representation Phosphosignatures Measured by Quantitative Targeted MS Capture Cellular States and Enable Large-scale Comparison of Drug-induced Phenotypes. Abelin JG1, Patel J1, Lu X1, Feeney CM1, Fagbami L1, Creech AL1, Hu R1, Lam D1, Davison D1, Pino L1, Qiao JW1, Kuhn E1, Officer A1, Li J2, Abbatiello S1, Subramanian A1, Sidman R2, Snyder E3, Carr SA1, Jaffe JD4.

View Details (opens in a new window)

Mol Cell Proteomics. 2014 Jul;13(7):1690-704. doi: 10.1074/mcp.M113.036392. Epub 2014 Apr 9. Ischemia in tumors induces early and sustained phosphorylation changes in stress kinase pathways but does not affect global protein levels. Mertins P1, Yang F2, Liu T2, Mani DR3, Petyuk VA2, Gillette MA3, Clauser KR3, Qiao JW3, Gritsenko MA2, Moore RJ2, Levine DA4, Townsend R5, Erdmann-Gilmore P5, Snider JE5, Davies SR5, Ruggles KV6, Fenyo D6, Kitchens RT5, Li S5, Olvera N4, Dao F4, Rodriguez H7, Chan DW8, Liebler D9, White F10, Rodland KD2, Mills GB11, Smith RD2, Paulovich AG12, Ellis M5, Carr SA1


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva
Internal Standard
light stable isotope peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 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+) 3.4 5.6 5.6 5 10.2 9 6 11.6 10.6 15 15 15
y12 (2+) 3.6 4.3 5.4 5 9.6 9.7 6.2 10.5 11.1 15 15 15
y4 (1+) 4.6 5.4 4.2 6.3 7 7 7.8 8.8 8.2 15 15 15
sum 2.5 3 4.4 4.6 7.6 7.6 5.2 8.2 8.8 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1426 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
LFDQAFGLPR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
28
Peptide End
37
CPTAC ID
CPTAC-1426
Peptide Molecular Mass
1,162.6135
Species
Homo sapiens (Human)
Assay Type
Direct ImmunoMRM
Matrix
depleted digested plasma
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Large-Scale Interlaboratory Study to Develop, Analytically Validate and Apply Highly Multiplexed, Quantitative Peptide Assays to Measure Cancer-Relevant Proteins in Plasma. Abbatiello SE, Schilling B, Mani DR, Zimmerman LJ, Hall SC, MacLean B, Albertolle M, Allen S, Burgess M, Cusack MP, Gosh M, Hedrick V, Held JM, Inerowicz HD, Jackson A, Keshishian H, Kinsinger CR, Lyssand J, Makowski L, Mesri M, Rodriguez H, Rudnick P, Sadowski P, Sedransk N, Shaddox K, Skates SJ, Kuhn E, Smith D, Whiteaker JR, Whitwell C, Zhang S, Borchers CH, Fisher SJ, Gibson BW, Liebler DC, MacCoss MJ, Neubert TA, Paulovich AG, Regnier FE, Tempst P, Carr SA. Mol Cell Proteomics. 2015 Sep;14(9):2357-74. doi: 10.1074/mcp.M114.047050. Epub 2015 Feb 18. PMID: 25693799


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
heavy stable isotope peptide (delta 8-10)
Peptide Standard Purity
>95%
Peptide Standard Label Type
15N
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 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+) 3.5 2 1.6 9.4 6 2.8 10 6.3 3.2 15 15 15
y6 (1+) 3.8 2.6 1.2 9.6 6.7 2.5 10.3 7.2 2.8 15 15 15
y5 (1+) 4.6 1.6 1.5 12.2 6.3 3.4 13 6.5 3.7 15 15 15
sum 2.9 1.9 1.1 10.2 6.2 2.7 10.6 6.5 2.9 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1430 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
QLSSGVSEIR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
80
Peptide End
89
CPTAC ID
CPTAC-1430
Peptide Molecular Mass
1,074.5669
Species
Homo sapiens (Human)
Assay Type
Direct ImmunoMRM
Matrix
depleted digested plasma
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Large-Scale Interlaboratory Study to Develop, Analytically Validate and Apply Highly Multiplexed, Quantitative Peptide Assays to Measure Cancer-Relevant Proteins in Plasma. Abbatiello SE, Schilling B, Mani DR, Zimmerman LJ, Hall SC, MacLean B, Albertolle M, Allen S, Burgess M, Cusack MP, Gosh M, Hedrick V, Held JM, Inerowicz HD, Jackson A, Keshishian H, Kinsinger CR, Lyssand J, Makowski L, Mesri M, Rodriguez H, Rudnick P, Sadowski P, Sedransk N, Shaddox K, Skates SJ, Kuhn E, Smith D, Whiteaker JR, Whitwell C, Zhang S, Borchers CH, Fisher SJ, Gibson BW, Liebler DC, MacCoss MJ, Neubert TA, Paulovich AG, Regnier FE, Tempst P, Carr SA. Mol Cell Proteomics. 2015 Sep;14(9):2357-74. doi: 10.1074/mcp.M114.047050. Epub 2015 Feb 18. PMID: 25693799


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
heavy stable isotope peptide (delta 8-10)
Peptide Standard Purity
>95%
Peptide Standard Label Type
15N
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 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 (1+) 4.3 4.5 3.3 7.1 6.2 5.3 8.3 7.7 6.2 15 15 15
y6 (1+) 3.2 4.1 1.5 5.1 5.1 3.5 6 6.5 3.8 15 15 15
y8 (1+) 2.9 4 2.6 5.5 5.4 5.6 6.2 6.7 6.2 15 15 15
sum 2.2 4 2.2 5.2 5.3 4.6 5.6 6.6 5.1 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1428 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
VSLDVNHFAPDELTVK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
97
Peptide End
112
CPTAC ID
CPTAC-1428
Peptide Molecular Mass
1,782.9152
Species
Homo sapiens (Human)
Assay Type
Direct ImmunoMRM
Matrix
depleted digested plasma
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Large-Scale Interlaboratory Study to Develop, Analytically Validate and Apply Highly Multiplexed, Quantitative Peptide Assays to Measure Cancer-Relevant Proteins in Plasma. Abbatiello SE, Schilling B, Mani DR, Zimmerman LJ, Hall SC, MacLean B, Albertolle M, Allen S, Burgess M, Cusack MP, Gosh M, Hedrick V, Held JM, Inerowicz HD, Jackson A, Keshishian H, Kinsinger CR, Lyssand J, Makowski L, Mesri M, Rodriguez H, Rudnick P, Sadowski P, Sedransk N, Shaddox K, Skates SJ, Kuhn E, Smith D, Whiteaker JR, Whitwell C, Zhang S, Borchers CH, Fisher SJ, Gibson BW, Liebler DC, MacCoss MJ, Neubert TA, Paulovich AG, Regnier FE, Tempst P, Carr SA. Mol Cell Proteomics. 2015 Sep;14(9):2357-74. doi: 10.1074/mcp.M114.047050. Epub 2015 Feb 18. PMID: 25693799


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
heavy stable isotope peptide (delta 8-10)
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 1.9um, 200A
Column Dimensions
0.075 x 150 mm
Flow Rate
200 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
y15 (2+) 5.5 7.3 5.5 15.1 9.3 7.3 16.1 11.8 9.1 15 15 15
y14 (2+) 3.7 7.4 6.6 13.6 8.6 5.8 14.1 11.3 8.8 15 15 15
y13 (2+) 8.5 7.5 6.8 15.7 10.7 6.5 17.9 13.1 9.4 15 15 15
sum 3.7 6.3 5.8 13.5 8.8 6 14 10.8 8.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-702 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
DGVVEITGK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
115
Peptide End
123
CPTAC ID
CPTAC-702
Peptide Molecular Mass
916.4866
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
15N
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

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+) 25 9 1.6 28.8 10.3 3.1 38.1 13.7 3.5 15 15 15
y5 (1+) 13.5 3.2 0.8 16.4 4 2.9 21.2 5.1 3 15 15 15
y6 (1+) 14.9 2.1 1.5 17.6 3.8 2.6 23.1 4.3 3 15 15 15
sum 13.4 1.8 1 16 3.5 2.5 20.9 3.9 2.7 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-970 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Modified Sequence
GPS[+80.0]WDPFR
Modification Type
Phospho (ST)
Protein - Site of Modification
15
Peptide - Site of Modification
3
Peptide Start
13
Peptide End
20
CPTAC ID
CPTAC-970
Peptide Molecular Mass
1,040.4117
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
synthetic peptide
Peptide Standard Purity
Crude
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
b5 (1+) 30.4 6.5 3.9 24.4 8.5 7.6 39 10.7 8.5 15 24 15
y4 (1+) 29.5 6.1 5.2 22.2 7.7 8.5 36.9 9.8 10 15 24 15
y5 (1+) 26.3 7 7.3 28.6 9.7 12 38.9 12 14 15 24 15
sum 28.1 6.2 5.1 24.2 7.2 8.5 37.1 9.5 9.9 15 24 15


Additional Resources and Comments


Assay Details for CPTAC-704 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
LFDQAFGLPR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
28
Peptide End
37
CPTAC ID
CPTAC-704
Peptide Molecular Mass
1,162.6135
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
15N
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

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+) 18.5 3.8 3.4 22.3 4.3 5.8 29 5.7 6.7 15 15 15
y6 (1+) 22.3 3.5 2.9 20.9 5.9 6.8 30.6 6.9 7.4 15 15 15
y8 (1+) 20.1 6 8.1 21.7 13.6 9.5 29.6 14.9 12.5 15 15 15
sum 13.1 2.9 3 18.1 7.6 6.2 22.3 8.1 6.9 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-703 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPB1
Peptide Sequence
VSLDVNHFAPDELTVK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
97
Peptide End
112
CPTAC ID
CPTAC-703
Peptide Molecular Mass
1,782.9152
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 K
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

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
y13 (2+) 29.1 16.5 7.2 36.9 20.4 11.5 47 26.2 13.6 15 15 15
y14 (2+) 18 15.6 11.4 26.1 20.8 18.4 31.7 26 21.6 15 15 15
y15 (2+) 15.1 8.9 14.9 18.3 13 17.9 23.7 15.8 23.3 15 15 15
sum 16.4 8.7 8.9 17.6 13.3 13.4 24.1 15.9 16.1 15 15 15


Additional Resources and Comments