Overview

  • Product nameAnti-KAT3B / p300 antibody [NM11]
    See all KAT3B / p300 primary antibodies
  • Description
    Mouse monoclonal [NM11] to KAT3B / p300
  • Tested applicationsChIP, ICC/IF, IHC-P, IP, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig, Mink (Mustela), Monkey
  • Immunogen

    E1A affinity purified p300 protein.

  • EpitopeAmino acids 2071 - 2091.
  • Positive control
    • Ls174T cell lysate. Tonsil tissue section.

Properties

Applications

Our Abpromise guarantee covers the use of ab3164 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent dilution.
IHC-P Use a concentration of 2 - 4 µg/ml.
IP Use a concentration of 2 µg/ml. (native and denatured, with protein A)
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 300 kDa (predicted molecular weight: 289 kDa).
Flow Cyt Use 1µg for 106 cells.

Target

  • FunctionFunctions as histone acetyltransferase and regulates transcription via chromatin remodeling. Acetylates all four core histones in nucleosomes. Histone acetylation gives an epigenetic tag for transcriptional activation. Mediates cAMP-gene regulation by binding specifically to phosphorylated CREB protein. Also functions as acetyltransferase for nonhistone targets. Acetylates 'Lys-131' of ALX1 and acts as its coactivator in the presence of CREBBP. Acetylates SIRT2 and is proposed to indirectly increase the transcriptional activity of TP53 through acetylation and subsequent attenuation of SIRT2 deacetylase function. Acetylates HDAC1 leading to its inactivation and modulation of transcription. Acts as a TFAP2A-mediated transcriptional coactivator in presence of CITED2. Plays a role as a coactivator of NEUROD1-dependent transcription of the secretin and p21 genes and controls terminal differentiation of cells in the intestinal epithelium. Promotes cardiac myocyte enlargement. Can also mediate transcriptional repression. Binds to and may be involved in the transforming capacity of the adenovirus E1A protein. In case of HIV-1 infection, it is recruited by the viral protein Tat. Regulates Tat's transactivating activity and may help inducing chromatin remodeling of proviral genes. Acetylates FOXO1 and enhances its transcriptional activity.
  • Involvement in diseaseNote=Defects in EP300 may play a role in epithelial cancer.
    Note=Chromosomal aberrations involving EP300 may be a cause of acute myeloid leukemias. Translocation t(8;22)(p11;q13) with KAT6A.
    Rubinstein-Taybi syndrome 2 (RSTS2) [MIM:613684]: A disorder characterized by craniofacial abnormalities, postnatal growth deficiency, broad thumbs, broad big toes, mental retardation and a propensity for development of malignancies. Some individuals with RSTS2 have less severe mental impairment, more severe microcephaly, and a greater degree of changes in facial bone structure than RSTS1 patients. Note=The disease is caused by mutations affecting the gene represented in this entry.
  • Sequence similaritiesContains 1 bromo domain.
    Contains 1 KIX domain.
    Contains 2 TAZ-type zinc fingers.
    Contains 1 ZZ-type zinc finger.
  • DomainThe CRD1 domain (cell cycle regulatory domain 1) mediates transcriptional repression of a subset of p300 responsive genes; it can be de-repressed by CDKN1A/p21WAF1 at least at some promoters. It conatins sumoylation and acetylation sites and the same lysine residues may be targeted for the respective modifications. It is proposed that deacetylation by SIRT1 allows sumoylation leading to suppressed activity.
  • Post-translational
    modifications
    Acetylated on Lys at up to 17 positions by intermolecular autocatalysis. Deacetylated in the transcriptional repression domain (CRD1) by SIRT1, preferentially at Lys-1020.
    Citrullinated at Arg-2142 by PADI4, which impairs methylation by CARM1 and promotes interaction with NCOA2/GRIP1.
    Methylated at Arg-580 and Arg-604 in the KIX domain by CARM1, which blocks association with CREB, inhibits CREB signaling and activates apoptotic response. Also methylated at Arg-2142 by CARM1, which impairs interaction with NCOA2/GRIP1.
    Sumoylated; sumoylation in the transcriptional repression domain (CRD1) mediates transcriptional repression. Desumoylated by SENP3 through the removal of SUMO2 and SUMO3.
    Probable target of ubiquitination by FBXO3, leading to rapid proteasome-dependent degradation.
    Phosphorylated by HIPK2 in a RUNX1-dependent manner. This phosphorylation that activates EP300 happens when RUNX1 is associated with DNA and CBFB. Phosphorylated by ROCK2 and this enhances its activity. Phosphorylation at Ser-89 by AMPK reduces interaction with nuclear receptors, such as PPARG.
  • Cellular localizationCytoplasm. Nucleus. In the presence of ALX1 relocalizes from the cytoplasm to the nucleus. Co-localizes with ROCK2 in the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • CREBBP/EP300 inhibitory protein 1 antibody
    • Cyclic AMP responsive enhancer binding protein antibody
    • E1A associated protein p300 antibody
    • E1A binding protein p300 antibody
    • E1A-associated protein p300 antibody
    • EC 2.3.1.48 antibody
    • EP300 antibody
    • EP300: E1A binding protein p300 antibody
    • EP300_HUMAN antibody
    • Histone acetyltransferase p300 antibody
    • KAT3B antibody
    • p300 HAT antibody
    • RB and P300 binding protein EID 1 antibody
    • Retinoblastoma protein associated protein antibody
    • RSTS2 antibody
    see all

Anti-KAT3B / p300 antibody [NM11] images

  • ICC/IF image of ab3164 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3164, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor© 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43æM.

  • ab3164 (4µg/ml) staining KAT3B in human bladder using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of muscle nuclei.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Overlay histogram showing HeLa cells stained with ab3164 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3164, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-KAT3B / p300 antibody [NM11] (ab3164)

This product has been referenced in:
  • Gao Y  et al. Expression of p300 and CBP is associated with poor prognosis in small cell lung cancer. Int J Clin Exp Pathol 7:760-7 (2014). IHC-P ; Human . Read more (PubMed: 24551300) »
  • Qiu W  et al. Sublytic C5b-9 triggers glomerular mesangial cell apoptosis via XAF1 gene activation mediated by p300-dependent IRF-1 acetylation. Cell Death Dis 5:e1176 (2014). Read more (PubMed: 24743731) »

See all 9 Publications for this product

Product Wall

Application ChIP
Detection step Semiquantitative PCR
Sample Mouse Cell lysate - nuclear (Hippocampal neurons)
Specification Hippocampal neurons
Negative control IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37% formaldehyde
Positive control Pol II
Username

Grant Corbett

Verified customer

Submitted Jul 24 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (kidney cell line RCC4)
Total protein in input 100 µg
Specification kidney cell line RCC4
Immuno-precipitation step Protein A
Username

Abcam user community

Verified customer

Submitted Nov 09 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293A)
Loading amount 50 µg
Specification HEK293A
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%
Username

Abcam user community

Verified customer

Submitted Aug 16 2006

Thank you for your enquiry. The epitope that ab3164 recognizes is amino acids 2071 - 2091. If you have any additional questions, please contact us again.

Thank you for your recent e-mail to my colleague Jennifer. She is away this week unexpectedly and I will do my best to help you. I have looked at your protocol and I think the problem is that p300 is a nuclear protein and hence you should be using ...

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Thank you for your email and I'm sorry to hear that you are experiencing difficulty with this antibody. I do need some details from you, however, to help determine what the problem may be. What was the lot number that you received (located on the vial)...

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This antibody recognizes p300 as well as CBP. p300 and CBP differ by three noncontiguous residues within the 21 amino acid epitope region, a difference that does not detectably affect the reactivity of NM11 MAb.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"