Biotin, a 244Da vitamin (Vitamin H) molecule, exhibits an extraordinary binding affinity for avidin (Ka=1015 M-1). Biotin and avidin interaction is rapid and once the bond is established it can survive up to 3M guanidine-hydrochloride and extremes of pH. Biotin-avidin bonds can only be reversed by denaturing the avidin protein molecule with 8M guanidine-hydrochloride at pH1.5 or by boiling in SDS Page Sample Loading Buffer.
Avidin is a glycoprotein with approximately 10% of its total mass coming from carbohydrates. Avidin has a molecular weight of 67kDa and contains four identical 128 amino acid subunits that each has a single biotin binding domain. Avidin is a basic protein with an isoelectric pH of 10-10.5 and is readily soluble in aqueous buffers containing a wide range of salt, pH (2-11), temperature and other laboratory agents. This wide range of tolerance makes avidin suitable for a wide variety of analytical applications. Avidin has extraordinary binding affinity for biotin (Ka=1015M-1).
The avidin in covalently coupled to the agarose resulting in minimal leaching and is stable over pH2-11.
The Avidin Resin is designed for the single step small and large scale affinity purification of proteins and antibodies with a biotin tag. The resin can also be used for immunoprecipitations using biotin labeled antibodies. Supplied as a 50% resin slurry.
Specific Binding and Elution Buffers are also available.
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- Avidin covalently coupled to ~6% cross linked agarose. Minimal Leaching
- Binding capacity 15-20µg biotin/ml resin
- Bead Structure: 6% cross-linked agarose
- Immunoprecipitation with biotinylated antibodies
- Pull down assays with biotinylated proteins
- Purification of biotinylated molecules, including: