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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.

SMAD4

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Available Assays

Overview Data source: UniProt

Official Gene Symbol Other Aliases
SMAD4 DPC4, MADH4
Sequence Length (AA) Molecular Weight (Da)
552 60439
Protein Name
Mothers against decapentaplegic homolog 4
Sources
UniProt
PhosphoSitePlus ®
GeneCards 		Human Protein Atlas

Protein Sequence hover to view complete sequence

10 20 30 40 50
MDNMSITNTP TSNDACLSIV HSLMCHRQGG ESETFAKRAI ESLVKKLKEK
60 70 80 90 100
KDELDSLITA ITTNGAHPSK CVTIQRTLDG RLQVAGRKGF PHVIYARLWR
110 120 130 140 150
WPDLHKNELK HVKYCQYAFD LKCDSVCVNP YHYERVVSPG IDLSGLTLQS
160 170 180 190 200
NAPSSMMVKD EYVHDFEGQP SLSTEGHSIQ TIQHPPSNRA STETYSTPAL
210 220 230 240 250
LAPSESNATS TANFPNIPVA STSQPASILG GSHSEGLLQI ASGPQPGQQQ
260 270 280 290 300
NGFTGQPATY HHNSTTTWTG SRTAPYTPNL PHHQNGHLQH HPPMPPHPGH
310 320 330 340 350
YWPVHNELAF QPPISNHPAP EYWCSIAYFE MDVQVGETFK VPSSCPIVTV
360 370 380 390 400
DGYVDPSGGD RFCLGQLSNV HRTEAIERAR LHIGKGVQLE CKGEGDVWVR
410 420 430 440 450
CLSDHAVFVQ SYYLDREAGR APGDAVHKIY PSAYIKVFDL RQCHRQMQQQ
460 470 480 490 500
AATAQAAAAA QAAAVAGNIP GPGSVGGIAP AISLSAAAGI GVDDLRRLCI
510 520 530 540 550
LRMSFVKGWG PDYPRQSIKE TPCWIEIHLH RALQLLDEVL HTMPIADPQP
552
LD

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-1268 Collapse assay details

Data source: Panorama

Official Gene Symbol
SMAD4
Peptide Sequence
GFPHVIYAR
Modification Type
unmodified
Protein - Site of Modification
N/A
Peptide - Site of Modification
N/A
Peptide Start
89
Peptide End
97
CPTAC ID
CPTAC-1268
Peptide Molecular Mass
1,058.5662
Species
Homo sapiens (Human)
Assay Type
Direct MRM or SRM
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 R
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

Assay Multiplexing Expand assay panel

Pacific Northwest National Laboratory-DirectMRM2

CPTAC-1219:
AURKB.
LPLAQVSAHPWVR 		CPTAC-1220:
CHUK.
LGTGGFGNVC[+57.0]LYQHR 		CPTAC-1221:
CHUK.
VWAEAVHYVSGLK
CPTAC-1222:
JUN.
NSDLLTSPDVGLLK 		CPTAC-1223:
MGEA5.
EIPVESIEEVSK 		CPTAC-1224:
PCNA.
DLSHIGDAVVISC[+57.0]AK
CPTAC-1225:
PCNA.
FSASGELGNGNIK 		CPTAC-1226:
VAV1.
LNPGDIVELTK 		CPTAC-1227:
VAV1.
YC[+57.0]SQVESASK
CPTAC-1228:
AURKB.
SNVQPTAAPGQK 		CPTAC-1229:
ADCY9.
TDAHFVDVIK 		CPTAC-1230:
ADCY9.
VIPQHQLSISPDIR
CPTAC-1231:
ARHGEF1.
ELVPPDTLHSLPK 		CPTAC-1232:
B2M.
VEHSDLSFSK 		CPTAC-1233:
B3GNT8.
NLLLVRPLGPQASIR
CPTAC-1234:
CALCOCO1.
GAQELAASSQQK 		CPTAC-1235:
CALCOCO1.
LQLEGQVTELR 		CPTAC-1236:
CCNB1.
TALGDIGNK
CPTAC-1237:
CDKN1B.
NLFGPVDHEELTR 		CPTAC-1238:
CDKN1B.
VSNGSPSLER 		CPTAC-1239:
C3.
TIYTPGSTVLYR
CPTAC-1240:
EP300.
LGTFLENR 		CPTAC-1241:
ESR1.
LLFAPNLLLDR 		CPTAC-1242:
FOXM1.
VLLAEEGIAPLSSAGPGK
CPTAC-1243:
GNAI1.
DSGVQAC[+57.0]FNR 		CPTAC-1244:
GNAI1.
IAQPNYIPTQQDVLR 		CPTAC-1245:
GNAI3.
IDFGEAAR
CPTAC-1246:
GNAI3.
ISQSNYIPTQQDVLR 		CPTAC-1247:
GRB10.
SQQDPAGPGLPAQSDR 		CPTAC-1248:
HDAC1.
YYAVNYPLR
CPTAC-1249:
IL18.
ISTLSC[+57.0]ENK 		CPTAC-1250:
IL18.
SDIIFFQR 		CPTAC-1251:
JAK1.
QLASALSYLEDK
CPTAC-1252:
JAK2.
LSDPGISITVLPK 		CPTAC-1253:
CDC25B.
LLGHSPVLR 		CPTAC-1254:
CDC25B.
TAVNLPLER
CPTAC-1255:
NCOA1.
LVQGGGLDVLSER 		CPTAC-1256:
NCOA3.
AVSLDSPVSVGSSPPVK 		CPTAC-1257:
NCOA3.
LLQNGNSPAEVAK
CPTAC-1258:
NEK2.
SQDSSPVLSELK 		CPTAC-1259:
NEK2.
YSDELNEIITR 		CPTAC-1260:
NFKB1.
LSPAPSK
CPTAC-1261:
OGT.
AFLDSLPDVK 		CPTAC-1262:
OGT.
GSVAEAEDC[+57.0]YNTALR 		CPTAC-1263:
PCP4.
AAVAIQSQFR
CPTAC-1264:
PLK1.
HINPVAASLIQK 		CPTAC-1265:
RANBP10.
LYPAVNQQETPLPR 		CPTAC-1266:
RANBP10.
VQGTVHC[+57.0]FPISAR
CPTAC-1267:
SKP2.
LSDPIVNTLAK 		CPTAC-1268:
SMAD4.
GFPHVIYAR 		CPTAC-1269:
SMAD4.
YC[+57.0]QYAFDLK
CPTAC-1270:
STAT5A.
ATIISEQQAK 		CPTAC-1271:
STAT5A.
HILYNEQR 		CPTAC-1272:
STAT1.
NLSFFLTPPC[+57.0]AR
CPTAC-1273:
STAT2.
EVLQSLPLTEIIR 		CPTAC-1274:
STAT2.
HYQLLTEENIPENPLR 		CPTAC-1275:
STAT3.
FPELNYQLK
CPTAC-1276:
STAT3.
GLSIEQLTTLAEK 		CPTAC-1277:
STAT4.
YLYPDIPK

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
y4 (1+) 38.5 4.7 10 41.5 8.8 9 56.6 10 13.5 15 15 15
y5 (1+) 18.7 8.7 5.2 24.9 9.7 7.4 31.1 13 9 15 15 15
y7 (1+) 23 7.5 9.2 25.9 8.9 10.7 34.6 11.6 14.1 15 15 15
sum 19 6 4.2 21.7 8.1 5.8 28.8 10.1 7.2 15 15 15

Selectivity

Data source: Panorama

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Slope of Curve Fit for cell line
fragment_ion 1 2 3 4 5 6
y5 (1+) 0.96 1.04 1.03 1.16 1.19 1.2
y7 (1+) 0.98 1.04 1.02 1.17 1.22 1.19
y4 (1+) 0.99 1.21 0.97 1.18 1.19 1.22
sum 0.97 1.08 1.01 1.17 1.19 1.21

Stability

Data source: Panorama

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Fragment ion / Transition control_intra_CV actual_temp_intra_CV frozen_intra_CV FTx1_intra_CV FTx2_intra_CV
y5 (1+) 8.6 4.5 7.8 3.7 14.6
y4 (1+) 7 6.9 3.5 7.4 12.4
y7 (1+) 12 6.7 6 6.8 2.7
sum 6.9 3.5 6.1 4.7 11.9
Fragment ion / Transition all_intra_CV all_inter_CV
y5 (1+) 7.3 7.3
y4 (1+) 7.4 7.9
y7 (1+) 6.8 10.3
sum 6.1 6

Endogenous

Data source: Panorama

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Fragment ion / Transition intra_CV inter_CV total_CV
y4 (1+) 43.4 46.8 63.8
y5 (1+) 64.2 107.1 124.9
y7 (1+) 45.2 44.6 63.5
sum 52.3 69.8 87.2

Additional Resources and Comments

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

Assay Details for CPTAC-1269 Collapse assay details

Data source: Panorama

Official Gene Symbol
SMAD4
Peptide Modified Sequence
YC[+57.0]QYAFDLK
Modification Type
Carbamidomethyl (C)
Protein - Site of Modification
115
Peptide - Site of Modification
2
Peptide Start
114
Peptide End
122
CPTAC ID
CPTAC-1269
Peptide Molecular Mass
1,206.5379
Species
Homo sapiens (Human)
Assay Type
Direct MRM or SRM
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

Assay Multiplexing Expand assay panel

Pacific Northwest National Laboratory-DirectMRM2

CPTAC-1219:
AURKB.
LPLAQVSAHPWVR 		CPTAC-1220:
CHUK.
LGTGGFGNVC[+57.0]LYQHR 		CPTAC-1221:
CHUK.
VWAEAVHYVSGLK
CPTAC-1222:
JUN.
NSDLLTSPDVGLLK 		CPTAC-1223:
MGEA5.
EIPVESIEEVSK 		CPTAC-1224:
PCNA.
DLSHIGDAVVISC[+57.0]AK
CPTAC-1225:
PCNA.
FSASGELGNGNIK 		CPTAC-1226:
VAV1.
LNPGDIVELTK 		CPTAC-1227:
VAV1.
YC[+57.0]SQVESASK
CPTAC-1228:
AURKB.
SNVQPTAAPGQK 		CPTAC-1229:
ADCY9.
TDAHFVDVIK 		CPTAC-1230:
ADCY9.
VIPQHQLSISPDIR
CPTAC-1231:
ARHGEF1.
ELVPPDTLHSLPK 		CPTAC-1232:
B2M.
VEHSDLSFSK 		CPTAC-1233:
B3GNT8.
NLLLVRPLGPQASIR
CPTAC-1234:
CALCOCO1.
GAQELAASSQQK 		CPTAC-1235:
CALCOCO1.
LQLEGQVTELR 		CPTAC-1236:
CCNB1.
TALGDIGNK
CPTAC-1237:
CDKN1B.
NLFGPVDHEELTR 		CPTAC-1238:
CDKN1B.
VSNGSPSLER 		CPTAC-1239:
C3.
TIYTPGSTVLYR
CPTAC-1240:
EP300.
LGTFLENR 		CPTAC-1241:
ESR1.
LLFAPNLLLDR 		CPTAC-1242:
FOXM1.
VLLAEEGIAPLSSAGPGK
CPTAC-1243:
GNAI1.
DSGVQAC[+57.0]FNR 		CPTAC-1244:
GNAI1.
IAQPNYIPTQQDVLR 		CPTAC-1245:
GNAI3.
IDFGEAAR
CPTAC-1246:
GNAI3.
ISQSNYIPTQQDVLR 		CPTAC-1247:
GRB10.
SQQDPAGPGLPAQSDR 		CPTAC-1248:
HDAC1.
YYAVNYPLR
CPTAC-1249:
IL18.
ISTLSC[+57.0]ENK 		CPTAC-1250:
IL18.
SDIIFFQR 		CPTAC-1251:
JAK1.
QLASALSYLEDK
CPTAC-1252:
JAK2.
LSDPGISITVLPK 		CPTAC-1253:
CDC25B.
LLGHSPVLR 		CPTAC-1254:
CDC25B.
TAVNLPLER
CPTAC-1255:
NCOA1.
LVQGGGLDVLSER 		CPTAC-1256:
NCOA3.
AVSLDSPVSVGSSPPVK 		CPTAC-1257:
NCOA3.
LLQNGNSPAEVAK
CPTAC-1258:
NEK2.
SQDSSPVLSELK 		CPTAC-1259:
NEK2.
YSDELNEIITR 		CPTAC-1260:
NFKB1.
LSPAPSK
CPTAC-1261:
OGT.
AFLDSLPDVK 		CPTAC-1262:
OGT.
GSVAEAEDC[+57.0]YNTALR 		CPTAC-1263:
PCP4.
AAVAIQSQFR
CPTAC-1264:
PLK1.
HINPVAASLIQK 		CPTAC-1265:
RANBP10.
LYPAVNQQETPLPR 		CPTAC-1266:
RANBP10.
VQGTVHC[+57.0]FPISAR
CPTAC-1267:
SKP2.
LSDPIVNTLAK 		CPTAC-1268:
SMAD4.
GFPHVIYAR 		CPTAC-1269:
SMAD4.
YC[+57.0]QYAFDLK
CPTAC-1270:
STAT5A.
ATIISEQQAK 		CPTAC-1271:
STAT5A.
HILYNEQR 		CPTAC-1272:
STAT1.
NLSFFLTPPC[+57.0]AR
CPTAC-1273:
STAT2.
EVLQSLPLTEIIR 		CPTAC-1274:
STAT2.
HYQLLTEENIPENPLR 		CPTAC-1275:
STAT3.
FPELNYQLK
CPTAC-1276:
STAT3.
GLSIEQLTTLAEK 		CPTAC-1277:
STAT4.
YLYPDIPK

Chromatograms

Data source: Panorama

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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
y3 (1+) 6.5 5.8 9.2 9.6 10.5 9.8 11.6 12 13.4 15 15 15
y4 (1+) 4.5 2.6 4 11.8 8.3 9.5 12.6 8.7 10.3 15 15 15
y5 (1+) 3.9 2 6.3 10.3 7.6 9.7 11 7.9 11.6 15 15 15
sum 2.9 2.5 5.3 9.7 8.1 9.3 10.1 8.5 10.7 15 15 15

Selectivity

Data source: Panorama

view larger image
Slope of Curve Fit for cell line
fragment_ion 1 2 3 4 5 6
y5 (1+) 8.74 9.19 9.24 9.25 9.32 10.15
y4 (1+) 8.14 9.09 8.54 8.31 9.22 9.44
y3 (1+) 9.63 9.48 10 9.84 10.71 11.05
sum 8.76 9.23 9.18 9.08 9.59 10.13

Stability

Data source: Panorama

view larger image
Fragment ion / Transition control_intra_CV actual_temp_intra_CV frozen_intra_CV FTx1_intra_CV FTx2_intra_CV
Fragment ion / Transition all_intra_CV all_inter_CV

Endogenous

Data source: Panorama

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Fragment ion / Transition intra_CV inter_CV total_CV
y3 (1+) 46.1 58.7 74.6
y4 (1+) 50.5 57.3 76.4
y5 (1+) 34.3 36.4 50
sum 22.3 36.2 42.5

Additional Resources and Comments

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