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 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 A190-218A used for WB at 0.2 µg/ml (1:5,000). Detection: Chemiluminescence with an exposure time of 30 seconds.