Product Information

Kit supplied with the following:

One (1) vial containing 50μL of fully biotinylated MBP-AviTag^TM fusion protein at 1mg/mL (50μg).

One (1) vial containing 100μL of unbiotinylated MBP-AviTag^TM fusion protein at 1mg/mL (100μg).

By comparing a known amount of a 100% biotinylated AviTag^TM fusion protein as a standard against a known amount of BirA-reacted protein-of-interest in an ELISA-type, multi-well dilution format (called BRTA for Biotinylation Reaction Titration Assay) it is possible to assess the extent of biotinylation achieved in your own BirA biotinylation reactions in a highly accurate and reproducible manner.

In addition, a mass shift between biotinylated and unbiotinylated fusion proteins visible on both SDS-PAGE and Western blot can be used to subjectively determine the amount of biotinylation that has occurred in a reaction by the observation of differences in band density in a mixed population of biotinylated vs. nonbiotinylated substrate.

The BIS-300 Kit contains the fully biotinylated standard used for the BRTA protocol and an unbiotinylated AviTag"d protein to react in a parallel to your BirA/protein biotinylation reaction as a positive control. A link to the BRTA protocol can be found here.

   Avidity带有AviTag标签的人类全长ORF克隆已经被构建在多种T7和CMV启动子表达系统,可以立即购买使用。AviTag标签蛋白可以被体内或体外的生物素连接酶生物素化，而且抗生物素蛋白或链霉亲和素可以专一地与生物素结合，正基于这两个反应，AviTag?技术可以应用于蛋白的固定，纯化和显影。 虽然Avidity是一个小公司，科学带来的好处及其AviTag已得到认可。目前， AviTag技术已经在22个国家的世界十大制药公司和研究人员中得到认可。  Avidity Avitag™ Technology Avidity develops and sells molecular affinity tools for connecting molecules. Our patented 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. AviTag™ sequence is GLNDIFEAQKIEWHE. Oriented on streptavidin-coated surfaces, this creates an ideal presentation for molecular binding interactions. The scientific benefits of Avidity's AviTag Technology have garnered notice. Currently, AviTag technology is licensed by seven of the world's top ten pharmaceutical companies and is used by researchers in 22 countries. Company History Avidity, LLC was founded in 1996 by Millard Cull, Larry Lansing, Dr. Ron Gill, and Dr. Mark Seville to develop peptide tags and molecular biology products based upon the extraordinary affinity of biotin for avidin or streptavidin. While working at Affymax as a researcher, Millard was involved in the discovery of the AviTag technology, saw its commercial potential, and obtained the exclusive rights from Affymax. He started Avidity as a reagent company that sublicenses the right to use the AviTag technology to other companies and sells products for use with the technology. AviTag History AviTag is a patented biotin-tagging technology discovered at Affymax by Dr. Peter Schatz from a Directed Evolution approach called "Peptides-on-Plasmids." The Directed Evolution approach identified a number of peptide sequences that can be biotinylated by the BirA enzyme of E. coli, and the best of these sequences was called the AviTag. E. coli enzyme BirA biotinylates the 15 amino acid AviTag twice as fast as its natural substrate, the Biotin Carboxyl Carrier Protein (BCCP). The enzymatic biotinylation of proteins using AviTag technology is preferred over chemical biotinylation of proteins because BirA enzyme gently, and highly specifically, labels the lysine residue of the AviTag. Prior to the AviTag, the smallest biotin-accepting domain was comprised of 75 amino acids.