1. Basic Information

Name:	GRB2_HUMAN
Accession#:	P62993
Description:	Growth factor receptor-bound protein 2
AA Number:	217
Sequence:	1 51 101 151 201    MEAIAKYDFK ATADDELSFK RGDILKVLNE ECDQNWYKAE LNGKDGFIPK NYIEMKPHPW FFGKIPRAKA EEMLSKQRHD GAFLIRESES APGDFSLSVK FGNDVQHFKV LRDGAGKYFL WVVKFNSLNE LVDYHRSTSV SRNQQIFLRD IEQVPQQPTY VQALFDFDPQ EDGELGFRRG DFIHVMDNSD PNWWKGACHG QTGMFPRNYV TPVNRNV

*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

NQQIFLR

 

NYVTPVNR

 

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.