A6561

Sigma-Aldrich

Monoclonal Anti-Biotin–Alkaline Phosphatase antibody produced in mouse

clone BN-34, purified immunoglobulin, buffered aqueous glycerol solution

Synonym(s):
Monoclonal Anti-Biotin
NACRES:
NA.46
Pricing and availability is not currently available.

Quality Level

biological source

mouse

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

BN-34, monoclonal

form

buffered aqueous glycerol solution

application(s)

dot blot: 1:10,000 using human IgG and biotin-monoclonal anti-human IgG
immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:20 using human tonsil and biotin-monoclonal anti-human IgG
indirect ELISA: 1:30,000 using human IgG and biotin-monoclonal anti-human IgG
western blot (chemiluminescent): 1:10,000 using human IgG and biotin-monoclonal anti-human IgG

isotype

IgG1

conjugate

alkaline phosphatase conjugate

shipped in

wet ice

storage temp.

2-8°C

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General description

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.
Monoclonal Anti-Biotin (mouse IgG1 isotype) is derived from the BN-34 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with biotin-keyhole limpet hemocyanin (KLH).

Immunogen

Biotin-KLH conjugate

Application

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.
Monoclonal Anti-Biotin-Alkaline Phosphatase antibody produced in mouse has been used in:
  • blotting (western), Southern and dot blots)
  • immunohistology
  • in situ nucleic acids hybridization
  • enzyme linked immunosorbent assay (ELISA)
  • fluorescent activated cell-sorting (FACS)
  • electron microscopy

Biochem/physiol Actions

Biotin is a carboxylase co-enzyme that regulates the metabolism of amino acids, glucose and fatty acids. This co-enzyme has been implicated in processes such as immune responses, cell division and fetal growth. Biotin may also be involved in the epigenetic modifications of chromatin.
The high binding affinity of biotin to egg white or bacteria-derived avidin has been exploited in the design of immunoassays and immunohistologic staining techniques. Conventional immunoassay methods are improved using monoclonal anti-biotin, which enhances the sensitivity of avidin-biotin immunoassays by adding a second layer of enzyme via the biotin specific monoclonal antibody.

Physical form

Solution in 0.05 M Tris buffer, pH 8.0, containing 1% bovine serum albumin, 1 mM MgCl2, 50% glycerol and 15 mM sodium azide.

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.

Personal Protective Equipment

dust mask type N95 (US),Eyeshields,Gloves

RIDADR

NONH for all modes of transport

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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The Journal of nutritional biochemistry, 14(12), 680-690 (2003-12-24)
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