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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Application Key:

• WB-Western Blot 
• IP-Immunoprecipitation 
• IHC-Immunohistochemistry 
• ChIP-Chromatin Immunoprecipitation 
• IF-Immunofluorescence 
• F-Flow Cytometry 
• E-P-ELISA-Peptide

Species 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 
• All-All Species Expected

Product Description

Designed for use with rabbit polyclonal and monoclonal antibodies, this affinity purified goat anti-rabbit IgG (heavy and light chain) antibody is conjugated to horseradish peroxidase(HRP) for chemiluminescent detection.  This product is thoroughly validated with CST primary antibodies and will work optimally with the CST western immunoblotting protocol, ensuring accurate and reproducible results.

Product Usage Information

Recommended Antibody Dilutions:

1:1000–1:3000

20X LumiGLO® Reagent and 20X Peroxide #7003 1:1K–1:3K

SignalFireTM ECL Reagent #6883 1:1K–1:3K

SignalFireTM Plus ECL Reagent #12630 1:5K-1:15K

SignalFireTM Elite ECL Reagent #12757 1:10K-1:20K

Storage

Supplied in 10mM 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 antibodies. This product is stable for 36 months when stored at -20C.

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.