Monoclonal Anti-Biotin antibody produced in mouse, clone BN-34, ascites fluid

Application

Applications in which this antibody has been used successfully, and the associated peer-reviewed papers, are given below.Western Blotting (1 paper)

Utilized in a variety of applications including blotting, immunocytochemistyr, insitu nucleic acid hybridization, ELISA, fluorescent activated cell-sorting (FACS) and electron microscopy.Titer is approximately 1:2000 by ELISA but should be optimized by the individual user for their application of choiceAn essential vitamin, biotin is required by living organisms and cells in culture. Avidin′s high affinity for biotin is often exploited in immuno-based assays. While standard assay methods using the avidin-biotin enzyme complex will suffice for most studies, there are occasions when enhanced sensitivity is needed to detect smaller amounts of antigen or localize low density antigens in histologic sections. Conventional immunoassay methods are improved by the use of Monoclonal Anti-Biotin which enhances the sensitivity of avidin-biotin immunoassays by bridging a second layer of avidin-biotin-enzyme complex. This antibody can be used in many other applications where biotin can be introduced as a target label.Monoclonal Anti-Biotin antibody produced in mouse is suitable for ELISA at a working dilution of 1:4000 and for western blotting.

Monoclonal Anti-Biotin antibody has been used in proximity ligation assay (PLA), cell surface biotinylation assay and western blotting.

The antibody may be used in amplification techniques, ELISA, immunoblotting, in situ nucleic acid hybridization, flow cytometry, image analysis or confocal microscopy.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 may participate in the replication and transcription of DNA. It acts as a coenzyme in the fatty acid biosynthesis, glyconeogenesis, and catabolism of several branched-chain amino acids and odd-chain fatty acids.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Biotin is a water-soluble vitamin. It is also called as vitamin B8. Biotin is a bicyclic compound that has a uredo ring and a tetrahydrothiophene ring.

Monoclonal Anti-Biotin recognizes the free biotin molecule and biotin conjugated to various immunoglobulins in ELISA and immunohistochemical techniques. Specificity was verified by using biotinylated goat antibodies reactive against human and rabbit antigens coated on microwell plates.

The antibody recognizes free biotin and biocytin using competitive ELISA. Using indirect ELISA or immuno­histo­chemical staining, the antibody also recognizes biotin conjugated to various immunoglobulins. Specificity is evaluated using biotinylated goat antibodies to human and rabbit antigens coated on multiwell plates.

Immunogen

Biotinylated keyhole limpet hemocyanin (KLH)

Biotin-KLH conjugate

Preparation Note

Developed in BALB/c mice immunized with biotinylated KLH. Mouse myeloma cells and splenocytes are fused together to form a BN-34 hybridoma.