KAT3B/p300 Antibody (RW105) - BSA Free

Immunocytochemistry/Immunofluorescence: KAT3B/p300 Antibody (RW105) [NB100-616] - HeLa cells were fixed in 4% paraformaldehyde for 10 min and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-KAT3B Antibody (RW105)) at 5ug/ml for 60 minutes at room temperature and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective.

Western Blot: KAT3B/p300 Antibody (RW105) [NB100-616] - p300 detected in a HeLa nuclear extract using NB 100-616 (1:250). ECL: 20 minute exposure.

Immunocytochemistry/Immunofluorescence: KAT3B/p300 Antibody (RW105) [NB100-616] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-KAT3B/p300 Antibody (RW105) NB100-616 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: KAT3B/p300 Antibody (RW105) [NB100-616] - An intracellular stain was performed on RAW 246.7 cells with KAT3B/p300 antibody (RW105) NB100-616PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin. Image using the PE format of this antibody.

Flow (Intracellular): KAT3B/p300 Antibody (RW105) [NB100-616] - An intracellular stain was performed on THP-1 cells with KAT3B/p300 (RW105) antibody NB100-616 (blue) and a matched isotype control NBP2-27287 (orange). Cells were fixed with 4% PFA and permeabilized with 0.1% Saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).

Flow Cytometry: KAT3B/p300 Antibody (RW105) [NB100-616] - An intracellular stain was performed on Raw264.7 cells with KAT3B/p300 Antibody (RW105) NB100-616 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

• Reactivity: Hu, Mu, Rt, Md, Pm
• Applications: WB, ChIP, EM, Flow, Flow-IC, ICC/IF, In vitro, IP
• Clone: RW105
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

KAT3B/p300 Antibody (RW105) - BSA Free Summary

• Immunogen: Fusion protein containing residues 1572-2371 of human KAT3B/p300. [UniProt# Q0947]
• Localization: Nuclear
• Specificity: This is specific for p300 protein. This recognizes residues 1921-2023.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: EP300
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation
  • Electron Microscopy
  • Flow (Intracellular) 1ug/mL
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunoprecipitation 1:10-1:500
  • In vitro assay
  • Western Blot 1:250-1:500
• Application Notes: This KAT3B/p300 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunoprecipitation and Western Blot. Use in In-vitro and Electron Microscopy reported in scientific literature (PMID 25728767) Use In vitro assay reported in scientific literature (PMID: 25728767).
• Control:

  HeLa Nuclear Cell Lysate

Reactivity Notes

Human, mouse, rat, monkey, and mink.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for KAT3B/p300 Antibody (RW105) - BSA Free

• 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

In the eukaryotic genome, chromatin dynamicity is crucial for regulation of processes requiring access to genetic information and is regulated by several different factors including post-translational modifications of histone/non-histone components, especially via acetylation process which is catalyzed by enzymes known as lysine acetyltransferases (KATs). Among different KATs, KAT3B/ p300 and its homologue KAT3A/CBP (CREB-binding protein) are extensively studied KATs with well known cellular functions. KAT3B/ p300 and KAT3A/CBP have been shown to directly interact with several different DNA-binding transcription factors which includes nuclear hormone receptors, CREB, c-Fos, C-Jun/v-Jun, c-Myb/v-Myb, TFIIB and MyoD. Moreover, both KAT3B/p300 and KAT3A/CBP have been shown to exert HAT (histone acetyltransferase) activity, capable of acetylating all four core histone particles in nucleosomes and to play a key role in activating chromatin for transcription. KAT3B/p300 is a potent transcriptional coactivator that has been shown to regulate various biological processes such as cellular proliferation, differentiation, cell cycle, apoptosis, and response to genetic damage, and KAT3B/ p300 dysfunction interfering with these processes implicates in the pathogenesis of several diseases such as cancer, cardiac diseases, diabetes mellitus, and HIV.

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 (NB100-616)(9)

Publications using NB100-616

• Dhar SS, Zhao D, Lin T et al. MLL4 Is Required to Maintain Broad H3K4me3 Peaks and Super-Enhancers at Tumor Suppressor Genes Mol. Cell Jun 7 2018 [PMID: 29861161] (ChIP, Mouse)
• He M, Zheng B, Zhang Y et al. KLF4 mediates the link between TGF-beta1-induced gene transcription and H3 acetylation in vascular smooth muscle cells. FASEB J 2015 Sep [PMID: 26082460]
• Yi P, Wang Z, Feng Q et al. Structure of a biologically active estrogen receptor-coactivator complex on DNA Mol. Cell 2015 Mar 19 [PMID: 25728767] (In-vitro, EM)
• Byun SW, Chang YJ, Chung IS et al. Helicobacter pylori decreases p27 expression through the delta opioid receptor-mediated inhibition of histone acetylation within the p27 promoter Cancer Lett 2012 Aug 3 [PMID: 22867947] (ChIP, Human)
• Bhattacharya S, Michels CL, Leung MK, Arany ZP, Kung AL, Livingston DM. Functional role of p35srj, a novel p300/CBP binding protein, during transactivation by HIF-1. Genes Dev;13(1):64-75. 1999 Jan 1. [PMID: 9887100]
• Nemethova M, Wintersberger E. Polyomavirus large T antigen binds the transcriptional coactivator protein p300. J Virol;73(2):1734-9. 1999 Feb. [PMID: 9882390] (WB, IP, Mouse)
• Eckner R et al. Interaction and functional collaboration of p300/CBP and bHLH proteins in muscle and B-cell differentiation. Genes Dev;1 (19):2478-90. 1996 Oct 1. [PMID: 8843199]
• Eckner R et al. Association of p300 and CBP with simian virus 40 large T antigen. Mol Cell Biol;16(7):3454-64. 1996 Jul. [PMID: 8668161] (IP, Mouse)
• Eckner R et al. Molecular cloning and functional analysis of the adenovirus E1A-associated 300-kD protein (p300) reveals a protein with properties of a transcriptional adaptor. Genes Dev;8(8):869-84. 1994 Apr 15. [PMID: 7523245] (WB, Human)

FAQs for KAT3B/p300 Antibody (NB100-616). (Showing 1 - 1 of 1 FAQs).

1. I'm in the market for a monoclonal antibody for p300/KAT3B for WB, that would react with mouse p300. In your inventory you have some listed that I saw meet these criteria. I'm interested in knowing more about the RW antibodies: RW105, RW109 and RW128. Do you know if in addition to WB, any of these can also be used in IP or ChIP?
   • Thank you for contacting Novus Biologicals regarding our p300 antibodies. The three clones you are interested in are all guaranteed to work in mouse, and in WB and IP. We haven't yet tested these in ChIP, but if you wanted to try them and let us know how those results turn out you actually qualify for our Innovators Reward Program.

Bioinformatics

• Gene Symbol: EP300