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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
STAT3 APRF
Sequence Length (AA) Molecular Weight (Da)
770 88068
Protein Name
Signal transducer and activator of transcription 3
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MAQWNQLQQL DTRYLEQLHQ LYSDSFPMEL RQFLAPWIES QDWAYAASKE
60 70 80 90 100
SHATLVFHNL LGEIDQQYSR FLQESNVLYQ HNLRRIKQFL QSRYLEKPME
110 120 130 140 150
IARIVARCLW EESRLLQTAA TAAQQGGQAN HPTAAVVTEK QQMLEQHLQD
160 170 180 190 200
VRKRVQDLEQ KMKVVENLQD DFDFNYKTLK SQGDMQDLNG NNQSVTRQKM
210 220 230 240 250
QQLEQMLTAL DQMRRSIVSE LAGLLSAMEY VQKTLTDEEL ADWKRRQQIA
260 270 280 290 300
CIGGPPNICL DRLENWITSL AESQLQTRQQ IKKLEELQQK VSYKGDPIVQ
310 320 330 340 350
HRPMLEERIV ELFRNLMKSA FVVERQPCMP MHPDRPLVIK TGVQFTTKVR
360 370 380 390 400
LLVKFPELNY QLKIKVCIDK DSGDVAALRG SRKFNILGTN TKVMNMEESN
410 420 430 440 450
NGSLSAEFKH LTLREQRCGN GGRANCDASL IVTEELHLIT FETEVYHQGL
460 470 480 490 500
KIDLETHSLP VVVISNICQM PNAWASILWY NMLTNNPKNV NFFTKPPIGT
510 520 530 540 550
WDQVAEVLSW QFSSTTKRGL SIEQLTTLAE KLLGPGVNYS GCQITWAKFC
560 570 580 590 600
KENMAGKGFS FWVWLDNIID LVKKYILALW NEGYIMGFIS KERERAILST
610 620 630 640 650
KPPGTFLLRF SESSKEGGVT FTWVEKDISG KTQIQSVEPY TKQQLNNMSF
660 670 680 690 700
AEIIMGYKIM DATNILVSPL VYLYPDIPKE EAFGKYCRPE SQEHPEADPG
710 720 730 740 750
SAAPYLKTKF ICVTPTTCSN TIDLPMSPRT LDSLMQFGNN GEGAEPSAGG
760 770
QFESLTFDME LTSECATSPM

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 non-CPTAC-6251 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Modified Sequence
YC[+57.021464]RPESQEHPEADPGSAAPY[+79.966331]LK
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
N/A
Peptide - Site of Modification
8
Peptide Start
686
Peptide End
707
CPTAC ID
non-CPTAC-6251
Peptide Molecular Mass
2,581.0945
Species
Human
Assay Type
Enrichment MRM
Enrichment Method
N/A
Matrix
SKBR3 cells
Submitting Laboratory
Cell Signaling Technology
Submitting Lab PI
Yiying Zhu

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Orbitrap Q Exactive (Thermo)
Internal Standard
Heavy isotope-labeled peptides
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N
LC
EASY-nLC II (Thermo)
Column Packing
self-packed
Column Dimensions
ID 100um, length 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
y4 (1+) 5.6 6.8 10.5 9.5 6.7 12.7 11 9.5 16.5 15 15 15
y13 (2+) 15.6 6.7 10.8 16.2 5.6 14.3 22.5 8.7 17.9 15 15 15
b9 (2+) 10.1 7.1 10.8 13.6 6.2 12.8 16.9 9.4 16.7 15 15 15
sum 5.7 6.7 10.5 8.1 6.2 12.9 9.9 9.1 16.6 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-5962 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Sequence
TGVQFTTK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
4
Peptide Start
341
Peptide End
348
CPTAC ID
CPTAC-5962
Peptide Molecular Mass
880.4654
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
N/A
Matrix
FFPE tumor tissue lysate pool
Submitting Laboratory
Fred Hutchinson Cancer Research Center
Submitting Lab PI
Amanda Paulovich

Publication

View Details (opens in a new window)

Targeted Mass Spectrometry Enables Multiplexed Quantification of Immunomodulatory Proteins in Clinical Biospecimens.Whiteaker JR, Lundeen RA, Zhao L, Schoenherr RM, Burian A, Huang D, Voytovich U, Wang T, Kennedy JJ, Ivey RG, Lin C, Murillo OD, Lorentzen TD, Thiagarajan M, Colantonio S, Caceres TW, Roberts RR, Knotts JG, Reading JJ, Kaczmarczyk JA, Richardson CW, Garcia-Buntley SS, Bocik W, Hewitt SM, Murray KE, Do N, Brophy M, Wilz SW, Yu H, Ajjarapu S, Boja E, Hiltke T, Rodriguez H, Paulovich AG.Front Immunol. 2021 Nov 11;12:765898. doi: 10.3389/fimmu.2021.765898. eCollection 2021.PMID: 34858420


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
QTRAP 5500
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Eksigent nanoLC 425
Column Packing
3 µm ChromXP C18EP, 120 Å
Column Dimensions
150 x 0.3 mm
Flow Rate
0.3 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
y3 (1+) 15.1 15.9 8.1 21.9 21.2 12.5 26.6 26.5 14.9 13 25 14
b6 (1+) 26.9 23.1 18.1 33.2 23.5 20.4 42.7 33 27.3 13 25 14
b5 (1+) 16.3 17.6 13.7 18.8 19.2 14.3 24.9 26 19.8 13 25 14
b4 (1+) 13.5 17.3 8.4 15.5 20.9 13.3 20.6 27.1 15.7 13 25 14
b3 (1+) 17.3 5.6 6 14.5 14.1 10.8 22.6 15.2 12.4 13 25 14
y4 (1+) 5.3 5.5 4.7 7.4 11.5 7.6 9.1 12.7 8.9 13 25 14
y5 (1+) 10.3 6.7 5.6 11.8 13.9 11.7 15.7 15.4 13 13 25 14
y6 (1+) 8.9 7.8 5.2 7.9 16 12 11.9 17.8 13.1 13 25 14
sum 8.1 3.8 4.2 10 13.2 10 12.9 13.7 10.8 13 25 14


Additional Resources and Comments


Assay Details for non-CPTAC-5440 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Modified Sequence
FIC[+57.021464]VTPTTC[+57.021464]SNTIDLPMSPR
Modification Type
Carbamidomethyl (C), Carbamidomethyl (C)
Protein - Site of Modification
713, 719
Peptide - Site of Modification
3, 9
Peptide Start
710
Peptide End
729
CPTAC ID
non-CPTAC-5440
Peptide Molecular Mass
2,309.0854
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
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
b3 (1+) 9.2 4.2 3.6 10.7 4.4 4.4 14.1 6.1 5.7 15 15 15
y5 (1+) 7 2.7 3.1 6.4 4.2 4.3 9.5 5 5.3 15 15 15
y6 (1+) 7.9 4.8 4.1 8.1 4.7 5.9 11.3 6.7 7.2 15 15 15
sum 3.8 2.1 2.9 4.2 3.3 4.3 5.7 3.9 5.2 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5713 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Modified Sequence
FIC[+57.021464]VTPTTC[+57.021464]SNTIDLPMS[+79.966331]PR
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
710
Peptide End
729
CPTAC ID
non-CPTAC-5713
Peptide Molecular Mass
2,389.0517
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
N/A
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus R
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
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
y6-98 (1+) 28.2 9.8 7.6 38.8 13.2 5.9 48 16.4 9.6 15 15 15
y5 (1+) 8.3 7.1 5.3 10.4 13.8 7.9 13.3 15.5 9.5 15 15 15
y5-98 (1+) 12.6 7.4 7.3 19.8 11.9 7.4 23.5 14 10.4 15 15 15
b3 (1+) 20.1 4.7 5.2 22.8 9.3 7.1 30.4 10.4 8.8 15 15 15
sum 9 4.5 4.8 11.6 10.2 5.8 14.7 11.1 7.5 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5439 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Modified Sequence
YC[+57.021464]RPESQEHPEADPGSAAPYLK
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
688
Peptide - Site of Modification
2
Peptide Start
686
Peptide End
707
CPTAC ID
non-CPTAC-5439
Peptide Molecular Mass
2,501.1281
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
RSLCnano and Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N
LC
RSLCnano
Column Packing
C18 PepMap 100
Column Dimensions
75 micron ID x 25 cm length
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
y9 (2+) 12.4 5 4 10.6 4.8 6 16.3 6.9 7.2 15 15 15
y4 (1+) 6.4 2.1 2 7.3 2.9 3.8 9.7 3.6 4.3 15 15 15
b13 (2+) 4.8 1.9 3.1 6.7 2.3 3.1 8.2 3 4.4 15 15 15
b9 (1+) 14.1 6.5 3.3 13.3 6.8 5.2 19.4 9.4 6.2 15 15 15
b8 (1+) 13.9 2.4 4.9 11.1 4.2 4.5 17.8 4.8 6.7 15 15 15
sum 3.9 1.3 2 5.1 1.7 3.1 6.4 2.1 3.7 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5714 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Modified Sequence
YC[+57.021464]RPESQEHPEADPGSAAPY[+79.966331]LK
Modification Type
Phospho (Y)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
686
Peptide End
707
CPTAC ID
non-CPTAC-5714
Peptide Molecular Mass
2,581.0945
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
N/A
Matrix
Cell Lysate
Submitting Laboratory
Moffitt Cancer Center
Submitting Lab PI
John Koomen

Publication

View Details (opens in a new window)

Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling. Whiteaker JR, Sharma K, Hoffman MA, Kuhn E, Zhao L, Cocco AR, Schoenherr RM, Kennedy JJ, Voytovich U, Lin C, Fang B, Bowers K, Whiteley G, Colantonio S, Bocik W, Roberts R, Hiltke T, Boja E, Rodriguez H, McCormick F, Holderfield M, Carr SA, Koomen JM, Paulovich AG. Cell Reports Methods doi: 10.1016/j.crmeth.2021.100015.


Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
Thermo Quantiva
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N
LC
Dionex Ultimate 3000 RSLCnano
Column Packing
Pepmap 100
Column Dimensions
75um x 25cm
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+) 2.2 2 1.2 2.7 1.4 1.6 3.5 2.4 2 15 15 15
b9 (1+) 5.5 3.2 2.3 4.5 3.3 3.1 7.1 4.6 3.9 15 15 15
b13 (2+) 2.6 3.2 0.9 4 3.5 3 4.8 4.7 3.1 15 15 15
b9 (2+) 5.3 2.9 2.4 7 3.5 2.4 8.8 4.5 3.4 15 15 15
sum 1.5 1.3 1 2.7 1.5 2.1 3.1 2 2.3 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1275 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Sequence
FPELNYQLK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
355
Peptide End
363
CPTAC ID
CPTAC-1275
Peptide Molecular Mass
1,150.6023
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Ovarian cancer tumor tissue lysate
Submitting Laboratory
Pacific Northwest National Laboratory
Submitting Lab PI
Tao Liu

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
TSQ Vantage
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Waters nanoACQUITY (Part Number 176016000)
Column Packing
Waters BEH C18, 1.7 um 130 Å (Part Number 186007485)
Column Dimensions
100 um x 100 mm
Flow Rate
0.5 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+) 20.6 9.3 7.6 19.6 10.8 9 28.4 14.3 11.8 15 15 15
y6 (1+) 19.1 6.7 6.1 18.3 9.9 9.1 26.5 12 11 15 15 15
y7 (1+) 15.6 5.3 6.9 18.7 8 10.6 24.4 9.6 12.6 15 15 15
sum 11.9 5.8 5.6 11.7 8.5 8.8 16.7 10.3 10.4 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1276 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Sequence
GLSIEQLTTLAEK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
519
Peptide End
531
CPTAC ID
CPTAC-1276
Peptide Molecular Mass
1,401.7715
Species
Homo sapiens (Human)
Assay Type
Direct MRM
Matrix
Ovarian cancer tumor tissue lysate
Submitting Laboratory
Pacific Northwest National Laboratory
Submitting Lab PI
Tao Liu

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
TSQ Vantage
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N at C-terminus K
LC
Waters nanoACQUITY (Part Number 176016000)
Column Packing
Waters BEH C18, 1.7 um 130 Å (Part Number 186007485)
Column Dimensions
100 um x 100 mm
Flow Rate
0.5 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
y8 (2+) 70.1 33.3 51.6 105.1 39.9 72.1 126.3 52 88.7 15 15 15
y9 (2+) 48.3 22.7 17.4 63.3 27.5 27 79.6 35.7 32.1 15 15 15
y9 (1+) 6.2 10 13.1 27.9 16.6 16.3 28.6 19.4 20.9 15 15 15
sum 7.1 10 13 25.9 16.1 16.3 26.9 19 20.8 15 15 15


Additional Resources and Comments


Assay Details for CPTAC-1751 Collapse assay details

Data source: Panorama

Official Gene Symbol
STAT3
Peptide Modified Sequence
YC[+57.0]RPESQEHPEADPGSAAPY[+80.0]LK
Modification Type
Carbamidomethyl (C), Phospho (Y)
Protein - Site of Modification
687, 705
Peptide - Site of Modification
2, 20
Peptide Start
686
Peptide End
707
CPTAC ID
CPTAC-1751
Peptide Molecular Mass
2,581.0945
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
N/A
Matrix
Ovarian cancer cell lysate
Submitting Laboratory
Pacific Northwest National Laboratory
Submitting Lab PI
Tao Liu

Assay Parameters Collapse assay parameters

Data source: Panorama

Instrument
TSQ Vantage
Internal Standard
peptide
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N
LC
Waters nanoACQUITY (Part Number 176016000)
Column Packing
Waters BEH C18, 1.7 um 130 Å (Part Number 186007485)
Column Dimensions
100 um x 100 mm
Flow Rate
0.5 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
b13 (2+) 24.7 9.1 10.2 26.1 13.1 12.3 35.9 16 16 15 15 15
b8 (1+) 11.7 11.7 11.8 11.3 7.8 13.1 16.3 14.1 17.6 15 15 15
b9 (1+) 15.6 7.7 10.6 16.3 10 9.8 22.6 12.6 14.4 15 15 15
sum 12.4 4.9 6.1 11.1 7.8 6.7 16.6 9.2 9.1 15 15 15


Additional Resources and Comments