Rabbits were immunized with HA (YPYDVPDYA) 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 HA-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-HA antibody A190-108B 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.