Anti p53

Immunofluorescence microscopy of HeLa cells using anti p53. Rockland's Protein A purified Mab anti p53 was used at a 1:100 dilution in 10% normal goat serum in PBS and reacted overnight at 4 C. After washes cells were incubated with a 1:500 dilution of AlexaFluor594 Goat a Mouse IgG diluted in normal goat serum for 1 h at room temperature. Personnel Communication. Kuldeep Patel, Loyola University.

Immunofluorescence of Mouse anti-p53 antibody. Tissue: human brain. Fixation: free-floating. Antigen retrieval: not required. Primary antibody: anti-human-p53 antibody at 1:500 for 1 h at RT. Co-stained with YFP and Sox2 antibodies. Localization: p53 is nuclear and cytoplasmic. Staining: p53 as precipitated blue with Cy5, YFP as precipitated green with Cy2, and Sox2 as precipitated red with Cy3. z‐stacks from confocal expressed as one composite focal plain.

Western blotting using anti-p53 Antibody. Lane 1: HeLa whole cell lysate (p/n W09-000-364), Lane 2: HeLa cytosol fraction, and Lane 3: HeLa nuclear extract (p/n W09-001-367). Load of 15 ug was separated by 10% SDS-PAGE and transferred to nitrocellulose membrane. The membrane was blocked with 3% milk/TBST for 1 h at room temperature followed by incubation with Rockland's Protein A purified Mab anti-p53 antibody overnight at 4 C diluted 1:1,500 in blocking solution. The membrane was washed 3X with TBST and then incubated with a 1:2,000 dilution of HRP Goat-a-Mouse IgG (p/n 610-103-121) diluted in blocking buffer for 1 h at room temperature. After final washes the proteins reactive on the membrane were detected using ECL. Other detection systems will yield similar results. Personnel Communication. Kuldeep Patel, Loyola University.