Monoclonal Anti-Biotin–Agarose antibody produced in mouse

Biotin is an essential vitamin required for cells in living organisms and in cultures. The high binding affinity of biotin to egg white avidin or bacteria-derived streptavidin has been used to design immunoassays for detecting antigen-antibody interactions. The use of monoclonal anti-biotin IgG can enhance the sensitivity of such avidin-biotin immunoassays. The monoclonal antibody functions by selectively enlarging and bridging the avidin-biotin-enzyme complexes, thereby increasing the signals from substrate conversion. Furthermore, this antibody can be used in many other applications where biotin may be introduced as target label.

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.

Biotin-KLH conjugate

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

May be substituted for avidin to decrease non-specific binding.

Monoclonal Anti-Biotin-Agarose antibody produced in mouse has been used in:

• pulldown assay
• coimmunoprecipitation 
• chromatin immunoprecipitation
• purification of biotinylated compounds and biotin-containing compounds

Mouse monoclonal anti-biotin antibody was used to identify by western blot S-nitrosylated proteins that had been labeled with a biotin moeity based on the biotin-switch method. No reactivity was visible on the blots when the biotin switch method was inhibited by treatment with 100 mM dithiothreitol showing that this antibody shows no unspecific cross reactivity. The mouse monoclonal anti-biotin antibody does cross react with Arabidopsis plant by western blot, no cross reactivity with unbiotinylated proteins.
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.

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.

Suspension in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide

Prepared using purified antibody coupled to cyanogen bromide activated agarose

Polyclonal anti-biotin antibodies were shown to react with in vivo biotinylated proteins but not monoclonal anti-biotin antibodies.

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