Anti-KAT3B / p300 antibody [3G230 / NM-11] - ChIP Grade (ab14984)

Overview

  • Product nameAnti-KAT3B / p300 antibody [3G230 / NM-11] - ChIP GradeSee all KAT3B / p300 primary antibodies ...
  • Description
    Mouse monoclonal [3G230 / NM-11] to KAT3B / p300 - ChIP Grade
  • Tested applicationsIP, IHC-P, ICC/IF, IHC-FoFr, WB, ChIP, IHC-Frmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Recombinant full length protein (Human).

  • Positive control
    • This antibody gave a positive signal in HeLa and Jurkat whole cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab14984 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
IP Use at an assay dependent dilution.
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 21705428
WB Use a concentration of 1 µg/ml. Detects a band of approximately 300 kDa (predicted molecular weight: 300 kDa).
ChIP Use 2µg for 106 cells.
IHC-Fr Use at an assay dependent concentration.

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
    • p300 HAT antibody
    • RB and P300 binding protein EID 1 antibody
    • Retinoblastoma protein associated protein antibody
    • RSTS2 antibody
    see all

Anti-KAT3B / p300 antibody [3G230 / NM-11] - ChIP Grade images

  • ChIP was performed on HeLa chromatin from 4 million cells using 8ug of ab14984. Positive control locus IRS2 shows enrichment over the negative control MyoD as expected.

    This data was provided by an anonymous collaborator.

  • All lanes : Anti-KAT3B / p300 antibody [3G230 / NM-11] - ChIP Grade (ab14984) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate (ab14844)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 300 kDa
    Observed band size : 300 kDa
    Additional bands at : 75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
    This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab14984 overnight at 4°C. Antibody binding was detected using an anti-mouse secondary antibody conjugated to HRP, and visualised using ECL development solution.

References for Anti-KAT3B / p300 antibody [3G230 / NM-11] - ChIP Grade (ab14984)

This product has been referenced in:
  • Nahid MA  et al. Regulation of TLR2-mediated tolerance and cross-tolerance through IRAK4 modulation by miR-132 and miR-212. J Immunol 190:1250-63 (2013). Read more (PubMed: 23264652) »
  • Liu Y  et al. Akt kinase targets the association of CBP with histone H3 to regulate the acetylation of lysine K18. FEBS Lett 587:847-53 (2013). Read more (PubMed: 23434580) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 10 cells
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris Gel)
Sample Zebrafish Tissue lysate - whole (whole embryo lysate, deyolked, 10 embryos)
Specification whole embryo lysate, deyolked, 10 embryos
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Maximilian Krause

Verified customer

Submitted Apr 28 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (HeLa)
Loading amount 50 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (4-15% gel)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 23 2013

Thank you for your enquiry regarding ab14984. This information can be found on the on-line product datasheet:

http://www.abcam.com/ab14984 Anti-KAT3B / p300 antibody [3G230] - ChIP Grade

The unit size is 25 µg and the concentr...

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Thank you for contacting us.

I am sorry to hear that the antibody is not working as expected.

As we discussed over the phone, the following suggestions might be of help in improving your results:

1) loadingmore protein (e...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vialeach of ab40011 & ab14984.

To check the status of the order...

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Thank you for contacting us. We have the following antibodies in our catalog that where tested in ChIP: for CBP: ab2832, ab10489, and ab10490 for p300: ab14984 None of the antibodies has been tested so far in zebrafish. If you ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (pancreatic cancer)
Specification pancreatic cancer
Fixative acetone+methanol 1:1
Permeabilization Yes - PBS + TritonX-100 0.025%
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (pancreatic cancer)
Loading amount 35 µg
Specification pancreatic cancer
Gel Running Conditions Non-reduced Denaturing (7.5%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Dr. Pawel Mazur

Verified customer

Submitted Jun 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Kidney)
Specification Kidney
Fixative Paraformaldehyde
Permeabilization Yes - .2% Triton X-100
Blocking step Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Dr. Jonathan Rud

Verified customer

Submitted Dec 29 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (Kidney)
Specification Kidney
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde soln/Abcam ChIP K
Detection step Semiquantitative PCR
Positive control Construct which contained the promoter of interest, also used positive control suggestions from the Abcam ChIP kit
Negative control Used non-specific primers and isotype control antibodies
Username

Dr. Jonathan Rud

Verified customer

Submitted Dec 23 2009

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