1. Basic Information

Name:LDHA_HUMAN
Accession#:P00338
Description:L-lactate dehydrogenase A chain
AA Number:332
Sequence:
1
51
101
151
201
251
301
   MATLKDQLIY NLLKEEQTPQ NKITVVGVGA VGMACAISIL MKDLADELAL
VDVIEDKLKG EMMDLQHGSL FLRTPKIVSG KDYNVTANSK LVIITAGARQ
QEGESRLNLV QRNVNIFKFI IPNVVKYSPN CKLLIVSNPV DILTYVAWKI
SGFPKNRVIG SGCNLDSARF RYLMGERLGV HPLSCHGWVL GEHGDSSVPV
WSGMNVAGVS LKTLHPDLGT DKDKEQWKEV HKQVVESAYE VIKLKGYTSW
AIGLSVADLA ESIMKNLRRV HPVSTMIKGL YGIKDDVFLS VPCILGQNGI
SDLVKVTLTS EEEARLKKSA DTLWGIQKEL QF
*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

VTLTSEEEAR

 

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.