Chickens 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. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support. Antibody concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
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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, chicken anti-V5 antibody A190-118A used for WB at 0.2 µg/ml (1:5,000). Detection: Chemiluminescence with an exposure time of 30 seconds.