Rabbit anti-V5 Tag Antibody Affinity Purified
Product Details
Specifications
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Prior to conjugation, immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4. Molar enzyme/antibody protein ratio is 4:1. Rabbits were immunized with a synthetic peptide representing amino acid residues 95 to 108 (GKPIPNPLLGLDST) of RNA polymerase alpha subunit of simion virus 5 conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support and conjugated to horseradish peroxidase (HRP).
Prior to conjugation, immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4. Molar enzyme/antibody protein ratio is 4:1. Rabbits were immunized with V5 (GKPIPNPLLGLDST) conjugated to KLH.
Antibody was affinity purified using the peptide immobilized on solid support. Rabbit anti-V5 affinity purified antibodies were coupled to agarose beads using a cyanogen bromide method. Final concentration is 0.1 mg antibody in 0.4 ml of 50% slurry containing 0.2 ml agarose.
Immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4, before coupling to agarose beads.
Additional Product Information
The V5 epitope tag is an epitope tag with a 14-residue sequence of GKPIPNPLLGLDST. Derived from the RNA polymerase alpha subunit of simian parainfluenza virus 5, V5 tags can help with detection of proteins in western blot, IP, or microscopic techniques. also especially small and are less likely to interfere with protein function.
Applications
Not all listed applications have been specifically tested by our laboratory.
All western blot analysis is performed using 5% Milk-TBST for blocking and as antibody diluent. Primary antibody is incubated overnight.
Western blots of cell lysates are performed using Goat anti-Rabbit IgG Heavy and Light Chain Antibody (Cat. No. A120-101P).
Western blots of immunoprecipitates are performed using Goat anti-Rabbit Light Chain HRP Conjugate (Cat. No. A120-113P) with 5% Normal Pig Serum (Cat. No. S100-020) added to the blocking buffer.
All western blot analysis is performed using 5% Milk-TBST for blocking and as antibody diluent. Primary antibody is incubated overnight.
Western blots of cell lysates are performed using Goat anti-Rabbit IgG Heavy and Light Chain Antibody (Cat. No. A120-101P).