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Kit supplied with the following:
By comparing a known amount of a 100% biotinylated AviTagTM 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.
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn