Antibody was affinity purified using an epitope specific to p53 immobilized on solid support. The epitope recognized by A300-247A maps to a region between residues 50 and 100 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 50 and 100
2 - 8°C
Detection of human P53 by western blot. Samples: Whole cell lysate (5, 15 and 50 µg) from HEK293T cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-P53 antibody A300-247A (lot A300-247A-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 A300-247A (lot A300-247A-2) used for IP at 3 µg per reaction. P53 was also immunoprecipitated by rabbit anti-P53 antibody A300-249A. For blotting immunoprecipitated P53, A300-247A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Flow Cytometrical Analysis of p53 in Jurkat Cells. Jurkat cells were fixed in 1.5% PFA, and permeabilized in 90% Methanol. 1 X 10^6 cells were stained with 1.5 µg anti-KLH IgG control or anti-p53 [A300-247A] and secondary FITC-conjugated goat anti-rabbit (in a 150µl reaction). anti-KLH control IgG (black); anti-p53 (red).
Localization of p53 Binding Sites by ChIP-sequencing. Chromatin from MCF-7 was immunoprecipitated with anti-p53 antibody A300-247A and analyzed by DNA sequencing. The figure illustrates the peak distribution of p53 binding within a 250 Kb region of chromosome 6 as detected using anti-p53 A300-247A. ChIP-seq validation performed by Active Motif, Carlsbad, CA.