hh Human p300 Antibody AF3789: R&D Systems

Human p300 Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human p300 in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human p300
    His2283-His2414
    Accession # Q09472
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    50 µg/mL
    See below
  • Immunohistochemistry
    5-25 µg/mL
    See below
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/5 x 106 cells
    See below
  • Immunocytochemistry
    1-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Chromatin Immunoprecipitation (ChIP)
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. The fos promoter was detected by standard PCR.
Immunocytochemistry
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.
Immunohistochemistry
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.
Detection of Human p300 by Simple WesternTM. 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: p300

The E1A-binding protein p300 is a histone acetyltransferase that also acts to acetylate other proteins, such as p53. p300 acts as a transcriptional coactivator that can serve as an adaptor molecule to bridge to transcriptional regulators. In addition, p300 binds to PCNA and may participate in chromatin remodeling.

  • Long Name:
    E1A-binding Protein p300
  • Entrez Gene IDs:
    2033 (Human); 328572 (Mouse)
  • Alternate Names:
    E1A binding protein p300; E1A-associated protein p300; E1A-binding protein, 300kD; EC 2.3.1; EC 2.3.1.48; EP300; histone acetyltransferase p300; KAT3B; p300 HAT; p300; RSTS2
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Metabolic Reprogramming, Autophagy, and Reactive Oxygen Species Are Necessary for Primordial Germ Cell Reprogramming into Pluripotency
    Authors: D Sainz de l, A Moratilla, V Aparicio, C Lorca, Y Alcaina, D Martín, MP De Miguel
    Oxid Med Cell Longev, 2017;2017(0):4745252.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
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