Biotin is considered as an essential nutrient, though it is know to be secreted by some of the intestinal bacteria. It is found in liver and brain tissues in minute quantities.
Biotin:avidin second antibody signal systems involve the conjugation of biotin to a secondary antibody. Once the secondary antibody binds to a target primary antibody in an immunochemical assay, the signal is developed by allowing avidin which has been conjugated to an enzyme or signal molecule to bind to the antibody associated biotin molecule.
Specificity and reactivity are determined by Ouchterlony Double Diffusion and immunoelectrophoresis versus the biotin-carrier protein complex. No reactivity is observed versus the carrier protein.
Immunogen
Biotin-KLH conjugate
Application
Anti-Biotin antibody produced in goat has been used in multi-well plate assay, to identify desthiobiotinylating (analog of biotin) proteins. It has also been used in Colourimetric enzyme immunoassay.
In some applications, localization of biotinylated probes with avidin produces high background levels. Anti-biotin reagents may be substituted for avidin to decrease non-specific binding.
Biochem/physiol Actions
Biotin serves as a cofactor for carboxylases, necessary for oxidative metabolism. Biotin absorption in small intestine is reduced on consumption of avidin rich food such as egg white. Thus, greatly decreasing its bioavailability, leading to egg white injury. Reduced levels of biotin is normally observed during pregnancy. Biotin deficiency might be congenital, with defective proteins involved in biotin homeostasis.
Physical form
Lyophilized from 0.01 M sodium phosphate, 15 mM sodium chloride, pH 7.2
Reconstitution
Reconstitute with 0.135 M sodium chloride.
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