antibody

Anti–BirA [6C4c7]

153488

monoclonal

mouse

nonconjugated

6C4c7

western blot, immunocytochemistry

153488

IF WB

BirA R118G

BirA antibody, bioR antibody, dhbB antibody, JW3941 antibody

E.coli

E.coli BirA is a well-characterized multifunctional protein that either catalyses the transfer of biotin to biotin carboxyl carrier protein (BCCP), a acetyl-CoA carboxylase subunit or, alternatively, binds to the biotin operator resulting in transcriptional repression of the biotin operon. A mutant BirA (R118G) shows loss of DNA binding ability and promiscuity of substrate biotinylation. These properties of BirA R118G were used to develop an in vivo proximity labelling technique that identifies potential interacting proteins (BioID). In the BioID method, a protein of interest is tagged with BirA R118G and expressed in live cells. Addition of the biotin to the culture medium results in biotinylation of proteins in vicinity of the bait. The biotinylated proteins can then be affinity purified with biotin binding proteins such as strepavidin or avidin even under harsh denaturing conditions and subsequently identified by mass spectrometry or immunoblot analysis.

Mouse

GST fused to E.coli BirA R118G

IgG1 kappa

Cells overexpressing an E.coli BirA construct

35

Sp2/0-Ag14

C57BL/6

Epitope Tags

This antibody recognises the C-terminal half residues 2-150 R118G of E.coli BirA. It has not been tested on other species. Gene Symbol: birA, bioR, dhbB, JW3941. Entrez Gene Number is NP_418404. UniProt Number: P06709.