Revision 3
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, E-P

REACTIVITY:

Rab

SENSITIVITY:

MW (kDa):

Source/Isotype:

Rabbit 

Product Information

Product Usage Information

Recommended Antibody Dilutions:

ELISA-peptide 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 2 mg/ml bovine serum albumin (BSA), and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) recognizes biotin attached to proteins, peptides, oligonucleotides and solid matrices.

Species Reactivity:

Rabbit

Source / Purification

Monoclonal antibody is produced by immunizing animals with a biotinylated protein.

Product Description

Anti-biotin (D5A7) Rabbit mAb is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. This product has been optimized to detect biotinylated primary antibodies. HRP conjugated antibodies do not require incubation with a secondary antibody. *Do not mix this antibody in solution with any Anti-rabbit antibody. Anti-rabbit antibodies will cross react with this antibody and could result in decreased activity of both Anti-rabbit and Anti-biotin (D5A7) Rabbit mAb.

Background

Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting E-P: Peptide ELISA (DELFIA)

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 3
#5571

Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate)

Western Blotting Image 1: Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) Expand Image
Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr) using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (Biotinylated) primary antibody and Anti-Biotin (D5A7) Rabbit mAb (HRP Conjugate) as secondary detection antibody.
Product Image 1: Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) Expand Image
Relative fluorescence of biotinylated and non-biotinylated peptides determined by #5571 Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) detection using #7003 LumiGLO® Reagent and 20X Peroxide reagents.
Product Image 2: Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) Expand Image
Chemiluminescent detection of phosphorylated and non-phosphorylated Met peptides with #4033 Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (Biotinylated) in conjunction with #5571 Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate).