1. Basic Information

Name:	STAT3_HUMAN
Accession#:	P40763
Description:	Signal transducer and activator of transcription 3
AA Number:	770
Sequence:	1 51 101 151 201 251 301 351 401 451 501 551 601 651 701 751    MAQWNQLQQL DTRYLEQLHQ LYSDSFPMEL RQFLAPWIES QDWAYAASKE SHATLVFHNL LGEIDQQYSR FLQESNVLYQ HNLRRIKQFL QSRYLEKPME IARIVARCLW EESRLLQTAA TAAQQGGQAN HPTAAVVTEK QQMLEQHLQD VRKRVQDLEQ KMKVVENLQD DFDFNYKTLK SQGDMQDLNG NNQSVTRQKM QQLEQMLTAL DQMRRSIVSE LAGLLSAMEY VQKTLTDEEL ADWKRRQQIA CIGGPPNICL DRLENWITSL AESQLQTRQQ IKKLEELQQK VSYKGDPIVQ HRPMLEERIV ELFRNLMKSA FVVERQPCMP MHPDRPLVIK TGVQFTTKVR LLVKFPELNY QLKIKVCIDK DSGDVAALRG SRKFNILGTN TKVMNMEESN NGSLSAEFKH LTLREQRCGN GGRANCDASL IVTEELHLIT FETEVYHQGL KIDLETHSLP VVVISNICQM PNAWASILWY NMLTNNPKNV NFFTKPPIGT WDQVAEVLSW QFSSTTKRGL SIEQLTTLAE KLLGPGVNYS GCQITWAKFC KENMAGKGFS FWVWLDNIID LVKKYILALW NEGYIMGFIS KERERAILST KPPGTFLLRF SESSKEGGVT FTWVEKDISG KTQIQSVEPY TKQQLNNMSF AEIIMGYKIM DATNILVSPL VYLYPDIPKE EAFGKYCRPE SQEHPEADPG SAAPYLKTKF ICVTPTTCSN TIDLPMSPRT LDSLMQFGNN GEGAEPSAGG QFESLTFDME LTSECATSPM

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

2. Protein Separation

Sample Preparation: 1 million cells from cell line K526 were lysed in an immuno-precipitation buffer containing 20 mM Tris-HCL (pH 7.6), 150 mM NaCl, 1mN EDTA, 0.1% NP-40, 100ug/ml PMSF, and 1mM sodium vanadate. Cell lysates were vortexed on ice for 30 minutes and spun to clear. The Supernatant was then incubated with protein A, and then with ABL1 specific antibody that has already been bound to protein A. Samples were incubated overnight, washed and then the beads were boiled in gel loading buffer and run for 1 hour on an SDS page gel (12% Bis-Tris). Resulting gel bands were then excised, reduced, alkylated and digested overnight with trypsin.

 

3. LC-MS/MS Data

TLTDEELADWK

    No LC-MS/MS for this peptide.

SAFVVER

 

FPELNYQLK

 

TQIQSVEPYTK

 

TGVQFTTK

 

4. LC-MRM Screening

Peptides screening: Peptides and transitions for analysis were selected from previous LC-MS/MS experiments. The results were screened for those with the highest specificity and the most intense transitions.