1. Basic Information

Name:	ACTB_HUMAN
Accession#:	P60709
Description:	Actin, cytoplasmic 1
AA Number:	375
Sequence:	1 51 101 151 201 251 301 351    MDDDIAALVV DNGSGMCKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIVTNWD DMEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL SLYASGRTTG IVMDSGDGVT HTVPIYEGYA LPHAILRLDL AGRDLTDYLM KILTERGYSF TTTAEREIVR DIKEKLCYVA LDFEQEMATA ASSSSLEKSY ELPDGQVITI GNERFRCPEA LFQPSFLGME SCGIHETTFN SIMKCDVDIR KDLYANTVLS GGTTMYPGIA DRMQKEITAL APSTMKIKII APPERKYSVW IGGSILASLS TFQQMWISKQ EYDESGPSIV HRKCF

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

2. Protein Separation

Sample Preparation: Cells from HCT116 were lysed in 8M urea/ 100mM ammonium bicarbonate buffer on ice. The equivalent of 100,000 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. 1/4 of the resulting digest was then analyzed on a Thermo Scientific TSQ mass spectrometer.

 

3. LC-MS/MS Data

GYSFTTTAER

 

AGFAGDDAPR

 

SYELPDGQVITIGNER

 

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.