Anti-KAT3B / p300 antibody [RW128] (ab185977)

Mouse monoclonal KAT3B / p300 antibody [RW128]. Validated in WB, IHC, ICC/IF and tested in Mouse, Human. Independently reviewed in 1 review(s). Immunogen corresponding to fusion protein.

Overview

  • Product name

    Anti-KAT3B / p300 antibody [RW128]
    See all KAT3B / p300 primary antibodies
  • Description

    Mouse monoclonal [RW128] to KAT3B / p300
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Fusion protein corresponding to KAT3B/ p300. GST-EP300 Fusion protein produced in bacteria (residues 1572-2371)

  • Positive control

    • ICC-IF: CACO2 cells
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab185977 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
ICC/IF Use a concentration of 5 - 10 µg/ml.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 250 kDa (predicted molecular weight: 264 kDa).
IHC-P Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Functions 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. Mediates acetylation of histone H3 at 'Lys-122' (H3K122ac), a modification that localizes at the surface of the histone octamer and stimulates transcription, possibly by promoting nucleosome instability. Mediates acetylation of histone H3 at 'Lys-27' (H3K27ac). Also functions as acetyltransferase for nonhistone targets. Acetylates 'Lys-131' of ALX1 and acts as its coactivator. 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. Acetylates BCL6 wich disrupts its ability to recruit histone deacetylases and hinders its transcriptional repressor activity. Participates in CLOCK or NPAS2-regulated rhythmic gene transcription; exhibits a circadian association with CLOCK or NPAS2, correlating with increase in PER1/2 mRNA and histone H3 acetylation on the PER1/2 promoter. Acetylates MTA1 at 'Lys-626' which is essential for its transcriptional coactivator activity (PubMed:10733570, PubMed:11430825, PubMed:11701890, PubMed:12402037, PubMed:12586840, PubMed:12929931, PubMed:14645221, PubMed:15186775, PubMed:15890677, PubMed:16617102, PubMed:16762839, PubMed:18722353, PubMed:18995842, PubMed:23415232, PubMed:23911289, PubMed:23934153, PubMed:8945521). Acetylates XBP1 isoform 2; acetylation increases protein stability of XBP1 isoform 2 and enhances its transcriptional activity (PubMed:20955178). Acetylates PCNA; acetylation promotes removal of chromatin-bound PCNA and its degradation during nucleotide excision repair (NER) (PubMed:24939902). Acetylates MEF2D.
  • Involvement in disease

    Defects in EP300 may play a role in epithelial cancer.
    Chromosomal aberrations involving EP300 may be a cause of acute myeloid leukemias. Translocation t(8;22)(p11;q13) with KAT6A.
    Rubinstein-Taybi syndrome 2
  • Sequence similarities

    Contains 1 bromo domain.
    Contains 1 CBP/p300-type HAT (histone acetyltransferase) domain.
    Contains 1 KIX domain.
    Contains 2 TAZ-type zinc fingers.
    Contains 1 ZZ-type zinc finger.
  • Domain

    The 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. Deacetylated by SIRT2, preferentially at Lys-418, Lys-423, Lys-1542, Lys-1546, Lys-1549, Lys-1699, Lys-1704 and Lys-1707.
    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 localization

    Cytoplasm. Nucleus. In the presence of ALX1 relocalizes from the cytoplasm to the nucleus. Colocalizes with ROCK2 in the nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • E1A associated protein p300 antibody
    • E1A binding protein p300 antibody
    • E1A-associated protein p300 antibody
    • EP300 antibody
    • EP300: E1A binding protein p300 antibody
    • EP300_HUMAN antibody
    • Histone acetyltransferase p300 antibody
    • KAT3B antibody
    • p300 HAT antibody
    • RSTS2 antibody
    see all

Images

  • All lanes : Anti-KAT3B / p300 antibody [RW128] (ab185977) at 1 µg/ml

    Lane 1 : Human colon tissue lysate - total protein (ab30051)
    Lane 2 : Colon (Mouse) Tissue Lysate
    Lane 3 : Colon (Rat) Tissue Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate
    Lane 7 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 8 : NIH 3T3 (Mouse) Whole Cell Lysate
    Lane 9 : Recombinant Human KAT3B / p300 protein (ab82235)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 5000 µg/ml

    Performed under reducing conditions.

    Predicted band size: 264 kDa
    Observed band size: 250 kDa
    why is the actual band size different from the predicted?


    Exposure time: 12 minutes
  • ab185977 stained CACO2 cells. The cells were 4% formaldehyde fixed for 10 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 min and then blocked in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to block non-specific protein-protein interactions. The cells were then incubated with the antibody ab185977 (10µg/ml) and ab202272 (pseudo-colored red) Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594) (1/250 dilution) overnight at +4°C. The secondary antibody (colored green) was ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed  used at a 1/1000 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • IHC image of KAT3B / p300 staining in Human normal colon formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab185977, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

References

ab185977 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
ChIP
Sample
Mouse Cell lysate - whole cell (Th17)
Negative control
mouse IgG
Specification
Th17
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Positive control
RNA polymerase II (RP2)

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Verified customer

Submitted Aug 22 2017

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