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Streptavidin-HRP #3999
Protocols
- 3999:
- ELISA-P, Western Blotting
Specificity / Sensitivity
Streptavidin has a remarkably high affinity for its natural ligand, biotin. The complex and irregular structure of the biotin-binding site makes it highly optimized for biotin binding and confers great specificity to the streptavidin-biotin complexes (4).
Source / Purification
Streptavidin-HRP is purified by gel filtration. The conjugate product is free from unconjugated streptavidin and HRP.
Western Blotting
Western blot analysis of extracts from HeLa cells, untreated or treated with Paclitaxel #9807, using Phospho-Histone H3 (Ser10) (D2C8) Rabbit mAb (Biotinylated) #3642 and Streptavidin-HRP #3999 for detection.
ELISA
The relationship between lysate protein concentration from untreated or serum and Calyculin A-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Antibody Pair #7207 and Streptavidin-HRP as a detection system is shown. NIH/3T3 cells were starved overnight and stimulated with serum for 15 minutes. Calyculin A #9902 was added and the cells were incubated at 37°C for 15 minutes and then lysed.
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.
- Updyke, T.V. and Nicolson, G.L. (1984) J Immunol Methods 73, 83-95.
- Buckie, J.W. and Cook, G.M. (1986) Anal Biochem 156, 463-72.
- Chaiet, L. and Wolf, F.J. (1964) Arch Biochem Biophys 106, 1-5.
- Reznik, G.O. et al. (1998) Proc Natl Acad Sci USA 95, 13525-30.
Application References
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- 7002 STOP Solution
- 9999 Nonfat Dry Milk
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
