1. Basic Information

Name:	RAF1_HUMAN
Accession#:	P04049
Description:	RAF proto-oncogene serine/threonine-protein kinase
AA Number:	648
Sequence:	1 51 101 151 201 251 301 351 401 451 501 551 601    MEHIQGAWKT ISNGFGFKDA VFDGSSCISP TIVQQFGYQR RASDDGKLTD PSKTSNTIRV FLPNKQRTVV NVRNGMSLHD CLMKALKVRG LQPECCAVFR LLHEHKGKKA RLDWNTDAAS LIGEELQVDF LDHVPLTTHN FARKTFLKLA FCDICQKFLL NGFRCQTCGY KFHEHCSTKV PTMCVDWSNI RQLLLFPNST IGDSGVPALP SLTMRRMRES VSRMPVSSQH RYSTPHAFTF NTSSPSSEGS LSQRQRSTST PNVHMVSTTL PVDSRMIEDA IRSHSESASP SALSSSPNNL SPTGWSQPKT PVPAQRERAP VSGTQEKNKI RPRGQRDSSY YWEIEASEVM LSTRIGSGSF GTVYKGKWHG DVAVKILKVV DPTPEQFQAF RNEVAVLRKT RHVNILLFMG YMTKDNLAIV TQWCEGSSLY KHLHVQETKF QMFQLIDIAR QTAQGMDYLH AKNIIHRDMK SNNIFLHEGL TVKIGDFGLA TVKSRWSGSQ QVEQPTGSVL WMAPEVIRMQ DNNPFSFQSD VYSYGIVLYE LMTGELPYSH INNRDQIIFM VGRGYASPDL SKLYKNCPKA MKRLVADCVK KVKEERPLFP QILSSIELLQ HSLPKINRSA SEPSLHRAAH TEDINACTLT TSPRLPVF

*Highlighted peptides (with yellow background) have developed assays.
*Green background amino acids are PTMs.

2. Protein Separation

Sample Preparation: Cells from Melanoma cancer cell lines were lysed in 8M urea/ 100mM ammonium bicarbonate buffer on ice. The equivalent of 200,000 cells were loaded onto a SDS gel. The protein of interest was excised and ingel digestion with trypsin was performed after reduction and alkylation with TCEP and IAA. 1/6 of the resulting digest was then analyzed on a Thermo Scientific TSQ mass spectrometer.

 

3. LC-MS/MS Data

HLHVQETK

    No LC-MS/MS for this peptide.

SASEPSLHR

    No LC-MS/MS for this peptide.

GYASPDLSK

    No LC-MS/MS for this peptide.

IGDFGLATVK

    No LC-MS/MS for this peptide.

APVSGTQEK

    No LC-MS/MS for this peptide.

IGSGSFGTVYK

    No LC-MS/MS for this peptide.

4. LC-MRM Screening

Peptides screening: Peptides and transitions for analysis were selected using a predictive peptide program. The results were screened for those with the highest specificity and the most intense transitions.