Secondary Antibodies
Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) #5571
Protocols
- 5571:
- ELISA Peptide*, Western Blotting
* Product-specific protocol.
Specificity / Sensitivity
Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) recognizes biotin attached to proteins, peptides, oligonucleotides and solide matrices.
Western Blotting
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.
ELISA-Peptide
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.
ELISA-Peptide
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).
Description
Anti-biotin (D5A7) Rabbit mAb is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups.
Applications
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.
Application References
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Companion Products
For Research Use Only. Not For Use In Diagnostic Procedures.
