1. Basic Information

Name:	HBB_HUMAN
Accession#:	P68871
Description:	Hemoglobin subunit beta
AA Number:	147
Sequence:	1 51 101    MVHLTPEEKS AVTALWGKVN VDEVGGEALG RLLVVYPWTQ RFFESFGDLS TPDAVMGNPK VKAHGKKVLG AFSDGLAHLD NLKGTFATLS ELHCDKLHVD PENFRLLGNV LVCVLAHHFG KEFTPPVQAA YQKVVAGVAN ALAHKYH

*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

LLVVYPWTQR

 

VLGAFSDGLAHLDNLK

 

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.