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Anti–BirA [6C4c7]

Info

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

There are 5 reference entries for this reagent.

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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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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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Inventor Information

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