Revision 1

#3999Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

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Product Information

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 2 mg/ml BSA, and 50% glycerol. Store at –20°C.

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).

Species Reactivity:

All Species Expected

Source / Purification

Streptavidin-HRP is purified by gel filtration. The conjugate product is free from unconjugated streptavidin and HRP.

Product 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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#3999

Streptavidin-HRP

Western Blotting Image 1: Streptavidin-HRP Expand Image
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.