What is biotin protein ligase?
Biotin-protein ligase (EC 6.3. 4.15) activates biotin to form biotinyl 5′ adenylate and transfers the biotin to biotin-accepting proteins. It also functions as a biotin operon repressor. The protein is encoded by the birA gene.
How does Avi-tag work?
Avidity Avitag™ Technology Avidity develops and sells molecular affinity tools for connecting molecules. Our AviTag™ technology employs a highly targeted enzymatic conjugation of a single biotin on a unique 15 amino acid peptide tag using the biotin ligase (BirA) from E. coli.
What biotin means?
Also known as vitamin H, biotin is one of the B complex vitamins that help the body convert food into energy. The word “biotin” comes from the ancient Greek word “biotos,” which means “life” or “sustenance.” B vitamins, and specifically biotin, help keep your skin, hair, eyes, liver, and nervous system healthy.
What is biotin biotinylation?
Biotinylation is the process of attaching biotin to proteins and other macromolecules. Biotinylation reagents are available for targeting specific functional groups or residues, including primary amines, sulfhydryls, carboxyls and carbohydrates.
What is an AVI-tag?
The Avi-tag is enzymatically biotinylated by the E. coli biotin ligase BirA. All R&D Systems Avi-tag Biotinylated Proteins undergo rigorous QC testing to ensure high bioactivity and lot-to-lot consistency.
How does the Bira biotin protein ligase work?
The BirA biotin-protein ligase adds d-biotin covalently to biotin-acceptor peptides/proteins via an ATP intermediate (biotinyl 5′-adenylate) in a highly efficient and targeted manner. This kit includes 300 µg of BirA enzyme for those who require larger scale reactions or more frequent use of the BirA biotinylation reaction in their work.
How is the AviTag amino acid sequence biotinylated?
Please Note: We strongly recommend the use of the AviTag™ amino acid sequence over other Biotinylation of Peptides sequences. The BirA enzyme biotinylates the AviTag™ sequence at a reaction rate 2x the natural substrate (BCCP) and as much as an order of magnitude or more over the other peptide sequences in use.
How is the biotin-avidin interaction used in chromatography?
The well known biotin-avidin/streptavidin interaction is often exploited for affinity chromatography or protein immobilization on surfaces or substrates. Protein detection via anti-biotin antibodies or avidin/streptavidin-reporter enzyme conjugates (-HRP, -alkaline phosphatase) or fluorescent probes becomes possible.
Which is an enzymatically biotinylated protein adds d-biotin?
The BirA biotin-protein ligase (EC 6.3.4.15) adds d-biotin covalently to biotin-acceptor peptides/proteins via an ATP intermediate (biotinyl 5′-adenylate) in a highly efficient and targeted manner. The downstream applications of enzymatically biotinylated proteins are varied, important and powerful.