KAT3B/p300 Antibody [Unconjugated]

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.

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.

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.

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 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.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, ChIP, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

KAT3B/p300 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human p300
  His2283-His2414
  Accession # Q09472
• Specificity: Detects human p300 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: EP300
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Immunocytochemistry 1-15 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Simple Western 50 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for KAT3B/p300 Antibody [Unconjugated]

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for KAT3B/p300 Antibody (AF3789)(1)

Publication using AF3789

• D Sainz de l, A Moratilla, V Aparicio, C Lorca, Y Alcaina, D Martín, MP De Miguel Metabolic Reprogramming, Autophagy, and Reactive Oxygen Species Are Necessary for Primordial Germ Cell Reprogramming into Pluripotency Oxid Med Cell Longev, 2017-07-05;2017(0):4745252. 2017-07-05 [PMID: 28757909] (ICC, Mouse)

Bioinformatics

• Gene Symbol: EP300
• Uniprot:
  • Q09472()