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A1559

Sigma-Aldrich

Monoclonal Anti-Biotin–Agarose antibody produced in mouse

clone BN-34, purified immunoglobulin, PBS suspension

Synonym(s):
Monoclonal Anti-Biotin
NACRES:
NA.46

biological source

mouse

Quality Level

recombinant

expressed in mouse cell line

conjugate

agarose conjugate

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

BN-34, monoclonal

form

PBS suspension

extent of labeling

2 mg antibody per mL resin

technique(s)

ELISA: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG1

capacity

0.15-0.3 μmol/mL, agarose binding capacity (biotin-ligand conjugate)

application(s)

research pathology

shipped in

wet ice

storage temp.

2-8°C

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General description

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.

Immunogen

Biotin-KLH conjugate

Application

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

Physical form

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

Preparation Note

Prepared using purified antibody coupled to cyanogen bromide activated agarose

Other Notes

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

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.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

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Certificate of Origin

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SDS

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