Description
Product Usage Information
The optimal dilution of the anti-biotin antibody should be determined by titration. However, a final dilution of 1:500 - 1:2000 should yield acceptable results for flow cytometry assays.
Storage
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
Protocol
Available protocols: Flow Cytometry (Fixed/Permeabilized), Flow Cytometry (Live)
Specificity / Sensitivity
Anti-biotin (D5A7) Rabbit Monoclonal Antibody (Alexa Fluor
Species Reactivity: All Species Expected
Source / Purification
Monoclonal antibody is produced by immunizing animals with biotinylated KLH.
Background
Biotin is a water-soluble B complex vitamin (vitamin H or B7) that serves as a coenzyme in various metabolic functions. It transfers carbon dioxide for five carboxylase enzymes and is involved in the TCA cycle and gluconeogenesis. In addition, all cells require biotin for cell proliferation, production of fatty acids, and metabolism of fats and amino acids (1). Biotin recycling and attachment to histone proteins is catalyzed by an enzyme named biotinidase. Histone biotinylation is involved in regulating telomere attrition and cellular senescence. Biotinidase deficiency is a rare genetic disorder that can cause neurological and cutaneous symptoms if left untreated (2). Biotin has a high affinity for its natural ligand, avidin. The complex and irregular structure of the biotin-binding site makes it highly optimized for biotin binding and confers great specificity to the avidin-biotin complexes (3). This remarkable affinity has been exploited for numerous immunoassays, including ELISA, Flow Cytometry, Immunofluorescence, In Situ Hybridization, Affinity Chromatography, and Immunohistochemistry. Anti-biotin (D5A7) Rabbit mAb #5597 can be used as an alternative to avidins in order to minimize background and maximize signal intensity.
