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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
Hspa5 Grp78
Sequence Length (AA) Molecular Weight (Da)
655 72422
Protein Name
Endoplasmic reticulum chaperone BiP
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MMKFTVVAAA LLLLGAVRAE EEDKKEDVGT VVGIDLGTTY SCVGVFKNGR
60 70 80 90 100
VEIIANDQGN RITPSYVAFT PEGERLIGDA AKNQLTSNPE NTVFDAKRLI
110 120 130 140 150
GRTWNDPSVQ QDIKFLPFKV VEKKTKPYIQ VDIGGGQTKT FAPEEISAMV
160 170 180 190 200
LTKMKETAEA YLGKKVTHAV VTVPAYFNDA QRQATKDAGT IAGLNVMRII
210 220 230 240 250
NEPTAAAIAY GLDKREGEKN ILVFDLGGGT FDVSLLTIDN GVFEVVATNG
260 270 280 290 300
DTHLGGEDFD QRVMEHFIKL YKKKTGKDVR KDNRAVQKLR REVEKAKRAL
310 320 330 340 350
SSQHQARIEI ESFFEGEDFS ETLTRAKFEE LNMDLFRSTM KPVQKVLEDS
360 370 380 390 400
DLKKSDIDEI VLVGGSTRIP KIQQLVKEFF NGKEPSRGIN PDEAVAYGAA
410 420 430 440 450
VQAGVLSGDQ DTGDLVLLDV CPLTLGIETV GGVMTKLIPR NTVVPTKKSQ
460 470 480 490 500
IFSTASDNQP TVTIKVYEGE RPLTKDNHLL GTFDLTGIPP APRGVPQIEV
510 520 530 540 550
TFEIDVNGIL RVTAEDKGTG NKNKITITND QNRLTPEEIE RMVNDAEKFA
560 570 580 590 600
EEDKKLKERI DTRNELESYA YSLKNQIGDK EKLGGKLSSE DKETMEKAVE
610 620 630 640 650
EKIEWLESHQ DADIEDFKAK KKELEEIVQP IISKLYGSGG PPPTGEEDTS
655
EKDEL

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 non-CPTAC-6098 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPA5
Peptide Sequence
ELEEIVQPIISK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
622
Peptide End
633
CPTAC ID
non-CPTAC-6098
Peptide Molecular Mass
1,396.7813
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Bovine Serum Albumin
Submitting Laboratory
McGill University
Submitting Lab PI
Christoph H. Borchers, Gerald Batist

Publication

View Details (opens in a new window)

mTORC1-driven protein translation correlates with clinical benefit of capivasertib within a genetically preselected cohort of PIK3CA-altered tumours. Constance A Sobsey, Bjoern C Froehlich, Georgia Mitsa, Sahar Ibrahim, Robert Popp, Rene P Zahedi, Elza C de Bruin, Christoph H Borchers, Gerald Batist


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Agilent 6495B QQQ-MS
Internal Standard
Standard Isotope labeled internal standard peptides with 13Cx, 15Ny (R+10 or K+8)
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Infinity 1290
Column Packing
commercial (Agilent ZORBAX Eclipse Plus C18)
Column Dimensions
4.6 mm × 150 mm, (5 µm)
Flow Rate
0.4 mL/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
y2 (1+) 11.2 4.3 3.1 6 5.1 4.6 12.7 6.7 5.5 16 16 17
y5 (1+) 14 4.5 4.2 9.4 4.1 4.4 16.9 6.1 6.1 16 16 17
y6 (1+) 10 3.3 4.3 8.9 3.4 4.9 13.4 4.7 6.5 16 16 17
y7 (1+) 11 4.9 4.2 6.3 4.8 4.8 12.7 6.9 6.4 16 16 17
y9 (1+) 9.1 3.8 5 7.2 4.1 5 11.6 5.6 7.1 16 16 17
sum 10.3 2.1 2.1 4 1.5 1.6 11 2.6 2.6 16 16 17


Additional Resources and Comments


Assay Details for CPTAC-525 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPA5
Peptide Sequence
ELEEIVQPIISK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
622
Peptide End
633
CPTAC ID
CPTAC-525
Peptide Molecular Mass
1,396.7813
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
cell line lysate pool
Submitting Laboratory
Seoul National University / Korea Institute of Science and Technology and FHCRC
Submitting Lab PI
Youngsoo Kim

Publication

View Details (opens in a new window)

Demonstrating the feasibility of large-scale development of standardized assays to quantify human proteins. Kennedy JJ, Abbatiello SE, Kim K, Yan P, Whiteaker JR, Lin C, Kim JS, Zhang Y, Wang X, Ivey RG, Zhao L, Min H, Lee Y, Yu MH, Yang EG, Lee C, Wang P, Rodriguez H, Kim Y, Carr SA, Paulovich AG. Nat Methods. 2014 Feb;11(2):149-55. doi: 10.1038/nmeth.2763. Epub 2013 Dec 8. PMID: 24317253


Response curves acquired at Seoul National University / Korea Institute of Science and Technology. Repeatability data acquired at Fred Hutchinson Cancer Research Center.


Response curves acquired at Seoul National University / Korea Institute of Science and Technology. Repeatability data acquired at Fred Hutchinson Cancer Research Center.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
5500 QTRAP (ABSCIEX)
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
nanoLC-Ultra 2D, cHiPLC-nanoflex (Eksigent)
Column Packing
ChromXP C18-CL, 3 um, 120A
Column Dimensions
150 x 0.075 mm
Flow Rate
500 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
y9 (1+) 4.8 6.5 5.9 18.5 12.8 7.3 19.1 14.4 9.4 15 25 15
y8 (1+) 15.9 8.1 9.2 22 14.3 14.4 27.1 16.4 17.1 15 25 15
y7 (1+) 8.8 5.7 4.4 19.2 11.5 7.8 21.1 12.8 9 15 25 15
sum 7.8 5.8 4.9 18.5 12 8.4 20.1 13.3 9.7 15 25 15


Additional Resources and Comments


Assay Details for CPTAC-524 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPA5
Peptide Sequence
SQIFSTASDNQPTVTIK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
448
Peptide End
464
CPTAC ID
CPTAC-524
Peptide Molecular Mass
1,835.9265
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
cell line lysate pool
Submitting Laboratory
Seoul National University / Korea Institute of Science and Technology and FHCRC
Submitting Lab PI
Youngsoo Kim

Publication

View Details (opens in a new window)

Demonstrating the feasibility of large-scale development of standardized assays to quantify human proteins. Kennedy JJ, Abbatiello SE, Kim K, Yan P, Whiteaker JR, Lin C, Kim JS, Zhang Y, Wang X, Ivey RG, Zhao L, Min H, Lee Y, Yu MH, Yang EG, Lee C, Wang P, Rodriguez H, Kim Y, Carr SA, Paulovich AG. Nat Methods. 2014 Feb;11(2):149-55. doi: 10.1038/nmeth.2763. Epub 2013 Dec 8. PMID: 24317253


Response curves acquired at Seoul National University / Korea Institute of Science and Technology. Repeatability data acquired at Fred Hutchinson Cancer Research Center.


Response curves acquired at Seoul National University / Korea Institute of Science and Technology. Repeatability data acquired at Fred Hutchinson Cancer Research Center.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
5500 QTRAP (ABSCIEX)
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C,15N
LC
nanoLC-Ultra 2D, cHiPLC-nanoflex (Eksigent)
Column Packing
ChromXP C18-CL, 3 um, 120A
Column Dimensions
150 x 0.075 mm
Flow Rate
500 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
y11 (1+) 23.9 9.7 5.6 25.5 11.2 6.7 34.9 14.8 8.7 15 25 15
y10 (1+) 12.1 7.4 4.6 13.1 8.1 6.2 17.8 11 7.7 15 25 15
y9 (1+) 25.5 9.8 4.7 27.8 9 14.8 37.7 13.3 15.5 15 25 15
sum 11.8 6.9 4 15 8.1 6.9 19.1 10.6 8 15 25 15


Additional Resources and Comments


Assay Details for non-CPTAC-2616 Collapse assay details

Data source: Panorama

Official Gene Symbol
HSPA5
Peptide Sequence
ITPSYVAFTPEGER
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
62
Peptide End
75
CPTAC ID
non-CPTAC-2616
Peptide Molecular Mass
1,565.7726
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
plasma
Submitting Laboratory
UVic-Genome BC Proteomics Centre
Submitting Lab PI
Christoph Borchers

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
6490 Triple Quad (Agilent)
Internal Standard
synthetic peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
1290 LC (Agilent)
Column Packing
Zorbax Eclipse Plus C18, 1.8 µm
Column Dimensions
2.1 x 150 mm
Flow Rate
400 µL/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+) 10.3 9.1 10.6 11 11.2 14.1 15.1 14.4 17.6 15 15 15
y9 (1+) 11 9.8 12.3 9.7 12.1 12.2 14.7 15.6 17.3 15 15 15
y12 (2+) 7.2 8.8 7.7 10.4 8.7 8.2 12.6 12.4 11.2 15 15 15
y5 (1+) 19.5 7.9 12.6 18.9 8.8 7.9 27.2 11.8 14.9 15 15 15
y8 (1+) 8.1 9.6 7.3 15.6 12.3 7.3 17.6 15.6 10.3 15 15 15
sum 4.3 6.2 6.8 9.1 7.1 7.5 10.1 9.4 10.1 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-3387 Collapse assay details

Data source: Panorama

Official Gene Symbol
Hspa5
Peptide Sequence
ITPSYVAFTPEGER
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
62
Peptide End
75
CPTAC ID
non-CPTAC-3387
Peptide Molecular Mass
1,565.7726
Species
Mus musculus (Mouse)
Assay Type
Direct MRM
Matrix
Plasma
Submitting Laboratory
UVic-Genome BC Proteomics Centre
Submitting Lab PI
Christoph Borchers

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Agilent 6490/6495 QQQ
Internal Standard
synthetic peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
1290 LC (Agilent)
Column Packing
Zorbax Eclipse Plus C18, 1.8 µm
Column Dimensions
2.1 x 150 mm
Flow Rate
400 µL/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
y12 (2+) 3.7 3.2 2.5 15.6 8.4 4.4 16 9 5.1 15 15 15
y5 (1+) 5.8 3.6 4.5 9.7 7.2 5.6 11.3 8 7.2 15 15 15
y7 (1+) 6.6 4.4 3.7 12.7 7.4 4.6 14.3 8.6 5.9 15 15 15
y8 (1+) 4.8 3.9 3.6 15.7 8.9 3.8 16.4 9.7 5.2 15 15 15
y9 (1+) 6.4 4.5 3.2 13.9 10.9 4.7 15.3 11.8 5.7 15 15 15
sum 2.3 1.7 1.7 13.9 7.9 3 14.1 8.1 3.4 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-3388 Collapse assay details

Data source: Panorama

Official Gene Symbol
Hspa5
Peptide Sequence
NQLTSNPENTVFDAK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
83
Peptide End
97
CPTAC ID
non-CPTAC-3388
Peptide Molecular Mass
1,676.8006
Species
Mus musculus (Mouse)
Assay Type
Direct MRM
Matrix
Plasma
Submitting Laboratory
UVic-Genome BC Proteomics Centre
Submitting Lab PI
Christoph Borchers

Publication

View Details (opens in a new window)

Molecular phenotyping of laboratory mouse strains using 500 multiple reaction monitoring mass spectrometry plasma assays. Michaud SA, Sinclair NJ, Petrošová H, Palmer AL, Pistawka AJ, Zhang S, Hardie DB, Mohammed Y Eshghi A1, Richard VR, Sickmann A, Borchers CH. Commun Biol. 2018 Jun 27;1:78. doi: 10.1038/s42003-018-0087-6. eCollection 2018.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Agilent 6490/6495 QQQ
Internal Standard
synthetic peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
1290 LC (Agilent)
Column Packing
Zorbax Eclipse Plus C18, 1.8 µm
Column Dimensions
2.1 x 150 mm
Flow Rate
400 µL/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
b3 (1+) 12.7 6.4 6.1 23.5 8.7 7.4 26.7 10.8 9.6 15 15 15
b2 (1+) 6.9 3.6 4.3 13.4 7.5 4.9 15.1 8.3 6.5 15 15 15
y2 (1+) 8.7 8.4 6.9 18.8 8.5 10.7 20.7 12 12.7 15 15 15
y3 (1+) 17.6 4.6 4.7 22.2 13.2 9 28.3 14 10.2 15 15 15
y9 (1+) 7.3 5.3 4.1 15.1 10.1 4.9 16.8 11.4 6.4 15 15 15
sum 3.2 2.1 2.9 14.3 8.1 3.5 14.7 8.4 4.5 15 15 15


Additional Resources and Comments