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

Overview

  • Product name
    Anti-KAT3B / p300 antibody [3G230 / NM-11] - ChIP Grade
    See all KAT3B / p300 primary antibodies
  • Description
    Mouse monoclonal [3G230 / NM-11] to KAT3B / p300 - ChIP Grade
  • Tested applications
    Suitable for: IP, 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.
  • General notes

    This antibody clone is manufactured by Abcam.

    Alternative versions available:

    Anti-KAT3B / p300 antibody (HRP) [3G230 / NM-11] (ab200186)

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 concentration.
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).

We recommend using 3% milk as the blocking agent in Western Blot.

ChIP Use 2µg for 106 cells.
IHC-Fr Use at an assay dependent concentration.

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

  • 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 nuclear extract 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 minutesThis 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.

Protocols

References

This product has been referenced in:
  • Yi Q  et al. Islet-1 induces the differentiation of mesenchymal stem cells into cardiomyocyte-like cells through the regulation of Gcn5 and DNMT-1. Mol Med Rep 15:2511-2520 (2017). WB, ChIP ; Mouse . Read more (PubMed: 28447752) »
  • Ni S  et al. CBX7 suppresses cell proliferation, migration, and invasion through the inhibition of PTEN/Akt signaling in pancreatic cancer. Oncotarget 8:8010-8021 (2017). ChIP ; Human . Read more (PubMed: 28030829) »

See all 15 Publications for this product

Customer reviews and Q&As

Application
ChIP
Sample
Mouse Cell lysate - nuclear (LBetaT2 Gonadotrope cell line)
Negative control
LbetaT2 chromatin immunoprecipitated with ab14984 and tested with POMC promoter primers, inactive in LBetaT2 cells LbetaT2 chromatin immunoprecipitated with unrelevant IgG and tested with Lhb promoter specific primers
Specification
LBetaT2 Gonadotrope cell line
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: Formaldehyde 1%
Positive control
LbetaT2 chromatin immunoprecipitated with ab14984 and tested with Lhb promoter specific primers (Mouillet JF J Biol Chem. 2004 Feb 27279(9):7832-9. Epub 2003 Dec 17.)
Username

Mr. David L'Hote

Verified customer

Submitted Oct 30 2017

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...

Read More

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

1-10 of 12 Abreviews or Q&A

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