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Overview Data source: UniProt

Official Gene Symbol Other Aliases
ARAF ARAF1,PKS,PKS2
Sequence Length (AA) Molecular Weight (Da)
606 67585
Protein Name
Serine/threonine-protein kinase A-Raf
Sources
UniProt
PhosphoSitePlus ®
GeneCards
Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MEPPRGPPAN GAEPSRAVGT VKVYLPNKQR TVVTVRDGMS VYDSLDKALK
60 70 80 90 100
VRGLNQDCCV VYRLIKGRKT VTAWDTAIAP LDGEELIVEV LEDVPLTMHN
110 120 130 140 150
FVRKTFFSLA FCDFCLKFLF HGFRCQTCGY KFHQHCSSKV PTVCVDMSTN
160 170 180 190 200
RQQFYHSVQD LSGGSRQHEA PSNRPLNELL TPQGPSPRTQ HCDPEHFPFP
210 220 230 240 250
APANAPLQRI RSTSTPNVHM VSTTAPMDSN LIQLTGQSFS TDAAGSRGGS
260 270 280 290 300
DGTPRGSPSP ASVSSGRKSP HSKSPAEQRE RKSLADDKKK VKNLGYRDSG
310 320 330 340 350
YYWEVPPSEV QLLKRIGTGS FGTVFRGRWH GDVAVKVLKV SQPTAEQAQA
360 370 380 390 400
FKNEMQVLRK TRHVNILLFM GFMTRPGFAI ITQWCEGSSL YHHLHVADTR
410 420 430 440 450
FDMVQLIDVA RQTAQGMDYL HAKNIIHRDL KSNNIFLHEG LTVKIGDFGL
460 470 480 490 500
ATVKTRWSGA QPLEQPSGSV LWMAAEVIRM QDPNPYSFQS DVYAYGVVLY
510 520 530 540 550
ELMTGSLPYS HIGCRDQIIF MVGRGYLSPD LSKISSNCPK AMRRLLSDCL
560 570 580 590 600
KFQREERPLF PQILATIELL QRSLPKIERS ASEPSLHRTQ ADELPACLLS
606
AARLVP

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
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Phosphorylation Acetylation Ubiquitylation Other

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Assay Details for CPTAC-1339 Collapse assay details

Data source: Panorama

Official Gene Symbol
ARAF
Peptide Modified Sequence
DS[+80.0]GYYWEVPPSEVQLLK
Modification Type
Phospho (ST)
Protein - Site of Modification
299
Peptide - Site of Modification
2
Peptide Start
298
Peptide End
314
CPTAC ID
CPTAC-1339
Peptide Molecular Mass
2,088.9445
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
Matrix
Digested Cell Lysate
Submitting Laboratory
Broad Institute
Submitting Lab PI
Steven A. Carr

Publication

View Details (opens in a new window)

Abelin, et al. Mol Cell Proteomics. 2016 May;15(5):1622-41. doi: 10.1074/mcp.M116.058354. Epub 2016 Feb 24. PMID:26912667


Assay Parameters Collapse assay parameters Test button response curves

Data source: Panorama

Instrument
Quantiva TSQ
Internal Standard
medium stable isotope peptide (delta 6 amu)
Peptide Standard Purity
Crude (~60%)
Peptide Standard Label Type
13C at C-terminus K
LC
Easy NanoLC1000
Column Packing
Reprosil C18, 3 um, 200A
Column Dimensions
0.075 x 100 mm
Flow Rate
300 nL/min

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


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+) 23.8 4.1 4.1 26.7 6.5 8.7 35.8 7.7 9.6 15 15 15
b8 (1+) 26.9 10.6 8.2 33.8 10 17.2 43.2 14.6 19.1 15 15 15
b8-98 (1+) 23.5 9.6 7.5 29 13.3 14.9 37.3 16.4 16.7 15 15 15
sum 14.7 4 4.2 14.8 4.9 9.7 20.9 6.3 10.6 15 15 15

Additional Resources and Comments


Assay Details for CPTAC-1343 Collapse assay details

Data source: Panorama

Official Gene Symbol
ARAF
Peptide Modified Sequence
QHEAPSNRPLNELLT[+80.0]PQGPS[+80.0]PR
Modification Type
Phospho (ST), Phospho (ST)
Protein - Site of Modification
181, 186
Peptide - Site of Modification
15, 20
Peptide Start
167
Peptide End
188
CPTAC ID
CPTAC-1343
Peptide Molecular Mass
2,597.1789
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
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 Test button response curves

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

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


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
y18 (2+) 16.1 11.7 9.7 15.6 13.6 19.6 22.4 17.9 21.9 13 13 15
y7-98 (1+) 8.4 7.1 4.4 9.2 7.7 14.1 12.5 10.5 14.8 15 15 15
y7 (1+) 6.5 5.3 6.4 7 6.2 10.2 9.6 8.2 12 15 15 15
sum 4.1 5.6 4.2 5.1 5.7 12.8 6.5 8 13.5 15 15 15

Additional Resources and Comments


Assay Details for CPTAC-1344 Collapse assay details

Data source: Panorama

Official Gene Symbol
ARAF
Peptide Modified Sequence
QHEAPSNRPLNELLT[+80.0]PQGPSPR
Modification Type
Phospho (ST)
Protein - Site of Modification
181
Peptide - Site of Modification
15
Peptide Start
167
Peptide End
188
CPTAC ID
CPTAC-1344
Peptide Molecular Mass
2,517.2126
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
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 Test button response curves

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

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


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
y18 (2+) 16.3 9.5 7.1 19.3 10.8 9.8 25.3 14.4 12.1 14 15 15
y7 (1+) 5.7 6.3 7.7 8.2 15.3 11.3 10 16.5 13.7 15 15 15
y8-98 (1+) 13.6 13.9 8.9 15.9 23.2 12.5 20.9 27 15.3 15 15 15
sum 5.8 6.4 5.5 8.1 11.8 9.1 10 13.4 10.6 15 15 15

Additional Resources and Comments


Assay Details for CPTAC-1345 Collapse assay details

Data source: Panorama

Official Gene Symbol
ARAF
Peptide Modified Sequence
QHEAPSNRPLNELLTPQGPS[+80.0]PR
Modification Type
Phospho (ST)
Protein - Site of Modification
186
Peptide - Site of Modification
20
Peptide Start
167
Peptide End
188
CPTAC ID
CPTAC-1345
Peptide Molecular Mass
2,517.2126
Species
Homo sapiens (Human)
Assay Type
Enrichment MRM
Enrichment Method
Fe(III)-NTA
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 Test button response curves

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

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


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
y18-98 (2+) 10 10.9 11.7 8.8 11.6 11.2 13.3 15.9 16.2 15 15 15
y18 (2+) 7.3 7.2 6.3 9.2 9 9.5 11.7 11.5 11.4 15 15 15
y7 (1+) 6 6.8 10.5 8.1 15.4 10 10.1 16.8 14.5 14 15 15
sum 6.3 5.9 7.6 6.9 9.7 8.7 9.3 11.4 11.6 15 15 15

Additional Resources and Comments


Assay Details for non-CPTAC-3958 Collapse assay details

Data source: Panorama

Official Gene Symbol
Araf
Peptide Sequence
IGDFGLATVK
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
443
Peptide End
452
CPTAC ID
non-CPTAC-3958
Peptide Molecular Mass
1,019.5651
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 Test button response curves

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 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

Assay Multiplexing Expand assay panel

Broad Institute-Biologist curated set of targets across a number of important cancer pathways


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+) 16.1 3.7 2.5 16 5.4 3.4 22.7 6.5 4.2 15 15 15
y8 (1+) 5.3 3.2 2.9 7.9 4.4 5.5 9.5 5.4 6.2 15 15 15
y9 (1+) 8.2 2.4 2.9 11.9 4 5.7 14.5 4.7 6.4 15 15 15
b3 (1+) 6.1 2.5 4.4 7.2 4.9 5.4 9.4 5.5 7 15 15 15
y6 (1+) 7.2 3.4 2.6 12.5 4.6 4.3 14.4 5.7 5 15 15 15
sum 2.7 1.1 1.3 7.4 3.3 4 7.9 3.5 4.2 15 15 15

Additional Resources and Comments