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 biotin.Antibody concentration was determined by extinction coefficient prior to conjugation: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
2 - 8°C
Detection of V5-tagged Protein by western blot. Samples: 200, 100, or 50 ng of E. coli whole cell lysate expressing a multi-tag fusion protein. Antibodies: Affinity-purified, biotin-conjugated, rabbit anti-V5 antibody A190-120B used for WB at 0.04 µg/ml (1:25,000). Detection: Streptavidin-HRP (0.1 µg/ml) and chemiluminescence with an exposure time of 10 seconds.