Antibody was affinity purified using an epitope specific to p53 immobilized on solid support. The epitope recognized by A300-249A maps to a region between residues 375 and the C-terminus (residue 393) of human tumor protein p53 using the numbering given in SwissProt entry P04637 (GeneID 7157). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
antigen NY-CO-13, BCC7, cellular tumor antigen p53, LFS1, mutant tumor protein 53, P53, p53 tumor suppressor, phosphoprotein p53, transformation-related protein 53, TRP53, tumor protein 53, Tumor suppressor p53, tumor supressor p53
between 375 and C-term
2 - 8°C
Detection of human p53 by western blot. Samples: Whole cell lysate (50 µg) from HeLa, HEK293T, and Jurkat cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-p53 antibody A300-249A (lot A300-249A-2) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human p53 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HEK293T cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-p53 antibody A300-249A (lot A300-249A-2) used for IP at 6 µg per reaction. p53 was also immunoprecipitated by a previous lot of this antibody (lot A300-249A-1). For blotting immunoprecipitated p53, A300-249A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 1 second.