Anti–BirA [6C4c7]
Invented at A*STAR Accelerate Technologies Pte Ltd
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- References (5)
- Inventor Info
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| Catalogue Number | 153488 |
| Applications | IF WB |
| Antigen/Gene or Protein Targets | BirA R118G |
| Synonyms | BirA antibody, bioR antibody, dhbB antibody, JW3941 antibody |
| Reactivity | E.coli |
| Relevance |
Monoclonal antibody to investigate the E.coli BirA protein. Background and Research Application E.coli BirA is a well-characterized multifunctional protein that either catalyses the transfer of biotin to biotin carboxyl carrier protein (BCCP), an acetyl-CoA carboxylase subunit or, alternatively, binds to the biotin operator resulting in transcriptional repression of the biotin operon. This protein also activates biotin to form biotinyl-5'-adenylate and transfers the biotin moiety to biotin-accepting proteins. 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 streptavidin or avidin even under harsh denaturing conditions and subsequently identified by mass spectrometry or immunoblot analysis. |
| Host | Mouse |
| Immunogen | GST fused to E.coli BirA R118G |
| Immunogen UniProt ID | P06709 |
| Positive Control | Cells overexpressing an E.coli BirA construct |
| Subclass | IgG1 kappa |
| Molecular Weight (kDa) | 35 |
| Myeloma Used | Sp2/0-Ag14 |
| Strain | C57BL/6 |
| Notes |
Production Details Purified using multi-step affinity chromatography with protein A. Storage Conditions Store at -20 degrees frozen. Avoid repeated freeze/thaw cycles. Points of Interest 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. Concentration 1mg/ml as standard |
| Research Area | Epitope Tags |
References: 5 entries
Roux et al. 2012. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells J Cell Biol. 196(6):801-10. PMID: 22412018.
Choi-Rhee et al. 2004. Protein Sci. 13(11):3043-50. PMID: 15459338.
Goodchild et al. 2004. Mislocalization to the nuclear envelope: an effect of the dystonia-causing torsinA mutation. Proc Natl Acad Sci U S A. 101(3):847-52. PMID: 14711988.
Streaker et al. 2003. Coupling of protein assembly and DNA binding: biotin repressor dimerization precedes biotin operator binding. J Mol Biol. 325(5):937-48. PMID: 12527300.
Kwon et al. 2000. Multiple disordered loops function in corepressor-induced dimerization of the biotin repressor. J Mol Biol. 304(5):821-33. PMID: 11124029.
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References: 5 entries
Roux et al. 2012. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells J Cell Biol. 196(6):801-10. PMID: 22412018.
Choi-Rhee et al. 2004. Protein Sci. 13(11):3043-50. PMID: 15459338.
Goodchild et al. 2004. Mislocalization to the nuclear envelope: an effect of the dystonia-causing torsinA mutation. Proc Natl Acad Sci U S A. 101(3):847-52. PMID: 14711988.
Streaker et al. 2003. Coupling of protein assembly and DNA binding: biotin repressor dimerization precedes biotin operator binding. J Mol Biol. 325(5):937-48. PMID: 12527300.
Kwon et al. 2000. Multiple disordered loops function in corepressor-induced dimerization of the biotin repressor. J Mol Biol. 304(5):821-33. PMID: 11124029.
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