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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
FOXO3 FKHRL1, FOXO3A
Sequence Length (AA) Molecular Weight (Da)
673 71277
Protein Name
Forkhead box protein O3
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MAEAPASPAP LSPLEVELDP EFEPQSRPRS CTWPLQRPEL QASPAKPSGE
60 70 80 90 100
TAADSMIPEE EDDEDDEDGG GRAGSAMAIG GGGGSGTLGS GLLLEDSARV
110 120 130 140 150
LAPGGQDPGS GPATAAGGLS GGTQALLQPQ QPLPPPQPGA AGGSGQPRKC
160 170 180 190 200
SSRRNAWGNL SYADLITRAI ESSPDKRLTL SQIYEWMVRC VPYFKDKGDS
210 220 230 240 250
NSSAGWKNSI RHNLSLHSRF MRVQNEGTGK SSWWIINPDG GKSGKAPRRR
260 270 280 290 300
AVSMDNSNKY TKSRGRAAKK KAALQTAPES ADDSPSQLSK WPGSPTSRSS
310 320 330 340 350
DELDAWTDFR SRTNSNASTV SGRLSPIMAS TELDEVQDDD APLSPMLYSS
360 370 380 390 400
SASLSPSVSK PCTVELPRLT DMAGTMNLND GLTENLMDDL LDNITLPPSQ
410 420 430 440 450
PSPTGGLMQR SSSFPYTTKG SGLGSPTSSF NSTVFGPSSL NSLRQSPMQT
460 470 480 490 500
IQENKPATFS SMSHYGNQTL QDLLTSDSLS HSDVMMTQSD PLMSQASTAV
510 520 530 540 550
SAQNSRRNVM LRNDPMMSFA AQPNQGSLVN QNLLHHQHQT QGALGGSRAL
560 570 580 590 600
SNSVSNMGLS ESSSLGSAKH QQQSPVSQSM QTLSDSLSGS SLYSTSANLP
610 620 630 640 650
VMGHEKFPSD LDLDMFNGSL ECDMESIIRS ELMDADGLDF NFDSLISTQN
660 670 673
VVGLNVGNFT GAKQASSQSW VPG

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-5836 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO3
Peptide Modified Sequence
AVS[+79.966331]MDNSNKYTK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
251
Peptide End
262
CPTAC ID
CPTAC-5836
Peptide Molecular Mass
1,436.6007
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
Matrix
Cell Lysate
Submitting Laboratory
Fred Hutchinson Cancer Research Center, H. Lee Moffitt Cancer Center & Research Institute, Broad Institute
Submitting Lab PI
Amanda Paulovich, John Koomen, Steven A. Carr

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
SCIEX 5500 / Thermo Quantiva
Internal Standard
Stable Isotope Labeled Standard
Peptide Standard Purity
>95%
Peptide Standard Label Type
13C and 15N K
LC
Dionex Ultimate 3000
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
y10-98 (2+) 7.9 10.3 7.4 22.5 16.2 16.3 23.8 19.2 17.9 16 15 16
y10 (2+) 8.2 11.2 8.4 21.2 16.6 17 22.7 20 19 16 15 16
sum 6.9 10.3 7.4 21.5 16.1 16.4 22.6 19.1 18 16 15 16


Additional Resources and Comments


Assay Details for non-CPTAC-5392 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO3
Peptide Sequence
AVSMDNSNK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
251
Peptide End
259
CPTAC ID
non-CPTAC-5392
Peptide Molecular Mass
964.4284
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 K
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
y5 (1+) 20.5 10.4 3.6 21.9 9.8 4.7 30 14.3 5.9 15 15 15
y6 (1+) 30.8 11.2 10.1 28.1 10.1 7 41.7 15.1 12.3 15 15 15
y7 (1+) 9.5 3.1 1.6 11.7 4.4 3 15.1 5.4 3.4 15 15 15
sum 8 3.8 1.6 10.6 4.3 3 13.3 5.7 3.4 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5740 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO3
Peptide Modified Sequence
AVS[+79.966331]MDNSNK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
251
Peptide End
259
CPTAC ID
non-CPTAC-5740
Peptide Molecular Mass
1,044.3947
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
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 K
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 (1+) 6.8 4.7 2.7 8.5 5.1 2.8 10.9 6.9 3.9 15 14 14
y5 (1+) 2.2 3.9 1.6 2.3 5.1 1.7 3.2 6.4 2.3 15 14 14
b3 (1+) 3.9 14 2.6 4.8 16.8 4.6 6.2 21.9 5.3 15 14 14
y7-98 (1+) 4.8 4.4 1.3 4.5 4.6 2.1 6.6 6.4 2.5 15 14 14
sum 2.5 3.2 0.9 2.9 4.2 1.1 3.8 5.3 1.4 15 14 14


Additional Resources and Comments


Assay Details for non-CPTAC-5394 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO3
Peptide Sequence
AVSMDNSNKYTK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
251
Peptide End
262
CPTAC ID
non-CPTAC-5394
Peptide Molecular Mass
1,356.6344
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 K
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
y8 (2+) 12.4 5.2 4 17.3 4.8 5.5 21.3 7.1 6.8 15 15 15
y9 (2+) 13.9 5.6 6.5 15.3 5.8 7.9 20.7 8.1 10.2 15 15 15
y10 (2+) 2.9 2.6 2.2 4 3.5 4.3 4.9 4.4 4.8 15 15 15
y3 (1+) 21.5 8.8 6.3 19.5 9 6.2 29 12.6 8.8 15 15 15
sum 2.7 2.7 2.1 3.9 3.4 4.3 4.7 4.3 4.8 15 15 15


Additional Resources and Comments


Assay Details for non-CPTAC-5739 Collapse assay details

Data source: Panorama

Official Gene Symbol
FOXO3
Peptide Modified Sequence
AVS[+79.966331]MDNSNKYTK
Modification Type
Phospho (ST)
Protein - Site of Modification
N/A
Peptide - Site of Modification
3
Peptide Start
251
Peptide End
262
CPTAC ID
non-CPTAC-5739
Peptide Molecular Mass
1,436.6007
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
IMAC
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 K
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
y8 (1+) 9 3.8 3.1 10.7 3.8 4.4 14 5.4 5.4 15 15 15
y7 (1+) 12.2 5.1 3.2 14 4.8 4.9 18.6 7 5.9 15 15 15
y3 (1+) 14.6 11.1 3.8 15.6 13.5 10.9 21.4 17.5 11.5 15 15 15
y11-98 (2+) 9.3 8.1 4.9 10.3 8.6 7.2 13.9 11.8 8.7 15 15 15
y10-98 (2+) 5.3 5.3 3.1 7.8 4.8 4.1 9.4 7.2 5.1 15 15 15
sum 5 3.9 2.1 5.5 4.5 4.9 7.4 6 5.3 15 15 15


Additional Resources and Comments