Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all p300 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human p300
His2283-His2414
Accession # Q09472

Specificity

Detects human p300 in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human p300 Antibody

Detection of Human p300 antibody by Western Blot.

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 antibody by Chromatin Immunoprecipitation.

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 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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 antibody by Simple WesternTM.

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.

p300 antibody in Human Colon Cancer Tissue by Immunohistochemistry (IHC-P).

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.

Applications for Human p300 Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: PMA and calcium ionomycin treated Jurkat human acute T cell leukemia cell line chromatin, fos promoter detected by standard PCR.

Immunocytochemistry

1-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer tissue

Simple Western

50 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

1 µg/mL
Sample: A431 human epithelial carcinoma cell line epidermoid carcinoma and HeLa human cervical epithelial carcinoma cell line cervical carcinoma cell lines

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

EP300

Entrez Gene IDs

2033 (Human); 328572 (Mouse)

Gene Symbol

EP300

UniProt

Q09472

Additional p300 Products

Product Documents for Human p300 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human p300 Antibody

For research use only

Related Research Areas

Notch Pathway
p53 Pathway

Citations for Human p300 Antibody

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Protocols

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