Antibody was affinity purified using an epitope specific to VCP immobilized on solid support. The epitope recognized by A300-589A maps to a region between residue 750 and the C-terminus (residue 806) of human Valosin-Containing Protein using the numbering given in entry NP_009057.1 (GeneID 7415). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
15S Mg(2+)-ATPase p97 subunit, CDC48, p97, TER ATPase, TERA, transitional endoplasmic reticulum ATPase, Valosin-containing protein, VCP
between 750 and C-term
2 - 8°C
Detection of human and mouse VCP by western blot. Samples: Whole cell lysate (15 µg) from HeLa, HEK293T, Jurkat, mouse TCMK-1, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-VCP antibody A300-589A (lot A300-589A-3) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 1 second.
Detection of human VCP by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-VCP antibody A300-589A (lot A300-589A-3) used for IP at 3 µg per reaction. VCP was also immunoprecipitated by rabbit anti-VCP antibody A300-588A. For blotting immunoprecipitated VCP, A300-589A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 1 second.