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Streptavidin-HRP #3999

Protocols

3999:

Western Blotting

Description

This Cell Signaling Technology product is useful for the detection of biotinylated proteins (1,2). Conjugation of horseradish peroxidase (HRP) to streptavidin is obtained by cross linking the amino groups on streptavidin with the carbohydrate groups on HRP.

Background

Streptavidin is a 53 kDa homotetramer isolated from Streptomyces avidinii for use in isolation and detection bioassays (3). Each streptavidin subunit forms high affinity non-covalent bonds with the vitamin biotin. Because of its strong non-covalent interaction with biotin, streptavidin can be used to detect and isolate biotinylated proteins (1,2).Chemiluminescent detection systems have emerged as the best all-around detection method for use with western blots and ELISA. These detection assays eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives. Streptavidin-HRP is used with biotinylated proteins and specific chemiluminescent substrates to generate light signal. Streptavidin-HRP conjugates have a very high turnover rate, coupling high sensitivity with short reaction times.

1. Updyke, T.V. and Nicolson, G.L. (1984) J Immunol Methods 73, 83-95.
2. Buckie, J.W. and Cook, G.M. (1986) Anal Biochem 156, 463-72.
3. Chaiet, L. and Wolf, F.J. (1964) Arch Biochem Biophys 106, 1-5.
4. Reznik, G.O. et al. (1998) Proc Natl Acad Sci USA 95, 13525-30.

Application References

• Liang, J. et al. (2011) Mol Cell Biol 31, 2577-90. Applications: Western Blotting

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For Research Use Only. Not For Use In Diagnostic Procedures.