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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.

CAST

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Available Assays

Overview Data source: UniProt

Official Gene Symbol Other Aliases
CAST N/A
Sequence Length (AA) Molecular Weight (Da)
708 76573
Protein Name
Calpastatin
Sources
UniProt
PhosphoSitePlus ®
GeneCards 		Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MNPTETKAIP VSQQMEGPHL PNKKKHKKQA VKTEPEKKSQ STKLSVVHEK
60 70 80 90 100
KSQEGKPKEH TEPKSLPKQA SDTGSNDAHN KKAVSRSAEQ QPSEKSTEPK
110 120 130 140 150
TKPQDMISAG GESVAGITAI SGKPGDKKKE KKSLTPAVPV ESKPDKPSGK
160 170 180 190 200
SGMDAALDDL IDTLGGPEET EEENTTYTGP EVSDPMSSTY IEELGKREVT
210 220 230 240 250
IPPKYRELLA KKEGITGPPA DSSKPIGPDD AIDALSSDFT CGSPTAAGKK
260 270 280 290 300
TEKEESTEVL KAQSAGTVRS AAPPQEKKRK VEKDTMSDQA LEALSASLGT
310 320 330 340 350
RQAEPELDLR SIKEVDEAKA KEEKLEKCGE DDETIPSEYR LKPATDKDGK
360 370 380 390 400
PLLPEPEEKP KPRSESELID ELSEDFDRSE CKEKPSKPTE KTEESKAAAP
410 420 430 440 450
APVSEAVCRT SMCSIQSAPP EPATLKGTVP DDAVEALADS LGKKEADPED
460 470 480 490 500
GKPVMDKVKE KAKEEDREKL GEKEETIPPD YRLEEVKDKD GKPLLPKESK
510 520 530 540 550
EQLPPMSEDF LLDALSEDFS GPQNASSLKF EDAKLAAAIS EVVSQTPAST
560 570 580 590 600
TQAGAPPRDT SQSDKDLDDA LDKLSDSLGQ RQPDPDENKP MEDKVKEKAK
610 620 630 640 650
AEHRDKLGER DDTIPPEYRH LLDDNGQDKP VKPPTKKSED SKKPADDQDP
660 670 680 690 700
IDALSGDLDS CPSTTETSQN TAKDKCKKAA SSSKAPKNGG KAKDSAKTTE
708
ETSKPKDD

Data source: UniProt

Protein Map Collapse protein map

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-1023 Collapse assay details

Data source: Panorama

Official Gene Symbol
CAST
Peptide Modified Sequence
S[+80.0]ESELIDELSEDFDR
Modification Type
Phospho (ST)
Protein - Site of Modification
364
Peptide - Site of Modification
1
Peptide Start
364
Peptide End
378
CPTAC ID
CPTAC-1023
Peptide Molecular Mass
1,862.7459
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
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

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Response Curves

Data source: Panorama

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Retrieving Data

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Repeatability

Data source: Panorama

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  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+) 13.9 73.4 102.4 55.8 59.9 69.8 57.5 94.7 123.9 7 13 9
y6 (1+) 27.6 25.1 19.6 34.9 47.6 37.5 44.5 53.8 42.3 10 18 12
y9 (1+) 27.3 21.2 13.1 33.1 47.6 16.7 42.9 52.1 21.2 10 18 12
sum 17.9 20.5 12 17.7 45.3 26.2 25.2 49.7 28.8 10 18 12

Additional Resources and Comments

Resources
Download SOPs
View Peptide in Panorama
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Comments

Assay Details for CPTAC-1024 Collapse assay details

Data source: Panorama

Official Gene Symbol
CAST
Peptide Modified Sequence
SES[+80.0]ELIDELSEDFDR
Modification Type
Phospho (ST)
Protein - Site of Modification
366
Peptide - Site of Modification
3
Peptide Start
364
Peptide End
378
CPTAC ID
CPTAC-1024
Peptide Molecular Mass
1,862.7459
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
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

view larger image
view larger image
view larger image

Response Curves

Data source: Panorama

view larger image
view larger image
view larger image

Retrieving Data

Loader

Repeatability

Data source: Panorama

view larger image
  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+) 38.3 70.6 58.7 32.1 60.6 95.5 50 93 112.1 7 13 9
y6 (1+) 47.1 18.2 18.2 47.7 53.8 28.2 67 56.8 33.6 10 18 12
y9 (1+) 34.4 19.2 16.2 41.4 55.5 27.5 53.8 58.7 31.9 10 18 12
sum 38 14 9.2 33.2 51.2 27.5 50.5 53.1 29 10 18 12

Additional Resources and Comments

Resources
Download SOPs
View Peptide in Panorama
View Protein in Panorama
Comments

Assay Details for non-CPTAC-3825 Collapse assay details

Data source: Panorama

Official Gene Symbol
Cast
Peptide Sequence
GVVPEDAVETLAGSLGTR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
505
Peptide End
522
CPTAC ID
non-CPTAC-3825
Peptide Molecular Mass
1,769.9159
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
>80%
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

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Response Curves

Data source: Panorama

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Retrieving Data

Loader

Repeatability

Data source: Panorama

view larger image
  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+) 8.2 3 2.7 12 10.5 12.2 14.5 10.9 12.5 15 15 15
sum 5.5 1.3 1.2 7.6 9.8 11.2 9.4 9.9 11.3 15 15 15
y15 (2+) 10.6 3.5 2.3 10.6 10.3 12.1 15 10.9 12.3 15 15 15
y3 (1+) 9.6 4.9 2.1 11.8 11.2 10.7 15.2 12.2 10.9 15 15 15
y7 (1+) 17.9 3.4 2.4 17.9 9.4 10.1 25.3 10 10.4 15 15 15

Additional Resources and Comments

Resources
Download SOPs
View Peptide in Panorama
View Protein in Panorama
Comments