Human p300 Antibody

Detection of Human p300 by Western Blot. Western blot shows nuclear extracts of A431 human epithelial carcinoma cell line epidermoid carcinoma and HeLa human cervical epithelial carcinoma cell line cervical carcinoma cell lines. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human p300 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3789) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for p300 at approximately 300 kDa (as indicated). This experiment was conducted using Immunoblot Buffer Group 1.

Detection of p300-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. p300/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human p300 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3789) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thefospromoter was detected by standard PCR.

p300 in HeLa Human Cell Line. p300 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human p300 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3789) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human p300 by Simple Western^TM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for p300 at approximately 308 kDa (as indicated) using 50 µg/mL of Goat Anti-Human p300 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3789) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

p300 in Human Colon Cancer Tissue. p300 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Goat Anti-Human p300 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3789) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.