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APPLICATIONS
Label Any Protein With Biotin
Avidity brings the power of the biotin-avidin interaction to a whole new range of biological applications. Using Biotin AviTag™ Technology, it is now possible to biotinylate just about any protein easily and efficiently at a single, unique site.
The Biotin AviTag sequence (Patent Nos. 5,932,433, 5,874,239 & 5,723,584) is a unique peptide, just 15 residues long, that is recognized by biotin ligase ( Schatz 1993). In the presence of ATP, the ligase specifically attaches biotin to the lysine residue in this sequence.
Using vectors developed by Avidity, the Biotin AviTag can be genetically fused to a much bigger protein. This feature effectively allows any protein that has been cloned to be tagged with a biotin molecule.
The simplicity and convenience of Biotin AviTag Technology is already being exploited by many researchers to rapidly immobilize a whole new range of target proteins for such purposes as drug screens and diagnostic assays.
A Superior Way of Labeling
The Biotin AviTag system, available exclusively from Avidity, affords several major advantages over the chemical labeling of proteins with biotin:
- Because the biotinylation is performed enzymatically, the reaction conditions are very gentle and the labeling is highly specific.
- Either in vivo or in vitro biotinylation of proteins is possible.
In addition, it should be noted that the Biotin AviTag has only one fifth of the bulk of alternative biotinylation tag sequences that are over 85 amino acid residues long; an important consideration if steric problems are to be minimized.
Available From Avidity
VERSATILE VECTORS FOR TAGGING
The Biotin AviTag can be easily introduced at either the N- or C- terminus or an external loop of any cloned target protein by using one of Avidity's unique vectors that come with the following features:- Multiple cloning sites.
- High copy number plasmid.
- A partitioning sequence for enhanced plasmid stability.
- A strong IPTG-inducible promoter.
SPECIAL STRAIN ENSURES BIOTINYLATION
Efficient biotinylation of Biotin AviTag-fused proteins can be achieved by expressing in Avidity's AVB101 E. coli B strain. AVB101 contains the pACYC184 ColE1 compatible plasmid that over-expresses the Biotin Ligase. Elevated levels of Biotin Ligase in the cells ensure complete biotinylation of Biotin AviTag-fused proteins.
THE BEST BIOTIN LIGASE
Fast and efficient in vitro biotinylation is achieved using Avidity's highly purified Biotin Ligase that comes complete with our exclusive reaction mix.
HELPFUL TECHNICAL SUPPORT
Expert technical support is readily available from the scientists who work with the Biotin AviTag system every day in the laboratory.
A Myriad of Applications
The availability and ease of use of Biotin AviTag Technology means that any protein can now be uniformly tethered to just about any surface including microtiter plates, chromatography media, biosensor chips and nitrocellulose/PVDF membranes.
The use of the Biotin AviTag is growing rapidly in various industrial and medical applications that demand an efficient and robust immobilization technology, including biosensors, diagnostics, proximity assays and drug screening.
Several major pharmaceutical and diagnostics corporations have already licensed the use of Biotin AviTags. If you, too, are interested in licensing Biotin AviTag Technology, please contact AVIDITY toll-free: 877.333.6063
ULTRA-SENSITIVE DETECTION TAGGED LUCIFERASE
Kikkoman Co., Japan, used a fully active Biotin AviTag-firefly luciferase fusion protein as an innovative marker enzyme for enzyme immunoassay (Tatsumi et al., 1996). Using this technique, they were able to detect as little as 2.5 pg/mL of test protein in a sandwich EIA. The Biotin AviTag-luciferase fusion provided 100,000 times more sensitive detection levels than other fusion constructs.
SUPERIOR PROTEIN PURIFICATION
Tucker and Grisshammer (1996) showed that neurotensin receptor with the Biotin AviTag fused to its C-terminus can be purified to homogeneity in just two steps. The crucial first step involved purification of the biotinylated protein on a monomeric avidin column. This yeilded essentially pure receptor with a remarkable 60-fold purification and 38% yield.
In side-by-side comparisons the purification protocol that employed the Biotin AviTag fusion was much more efficient than that provided by other affinity tags such as Hexa-His and Strep Tag.
EFFECTIVE IMMUNOLOGICAL SITINS
Altman et al. (1996) constructed tetrameric peptide-MHC complexes with which it was possible to develop an efficient immunological stain. The precise orientation afforded by the Biotin AviTag was essential to the success of this approach.
