Overview

  • Product nameAnti-KAT3B / p300 antibody - ChIP GradeSee all KAT3B / p300 primary antibodies ...
  • Description
    Mouse monoclonal to KAT3B / p300 - ChIP Grade
  • Tested applicationsIHC-P, ChIP, IP, WB, ICC/IF, Flow Cyt more details
  • Species reactivity
    Reacts with: Human, Recombinant Fragment
  • Immunogen

    Recombinant fragment: PPLQHHGQLA QPGALNPPMG YGPRMQQPSN QGQFLPQTQF PSQGMNVTNI PLAPSSGQAP VSQAQMSSSS CPVNSPIMPP GSQGSHIHCP QLPQPALHQN , corresponding to amino acids 731-831 of Human p300

Properties

Applications

Our Abpromise guarantee covers the use of ab54984 in the following tested applications.

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

Application Notes
IHC-P Use a concentration of 3 µg/ml.
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 - 5 µg/ml.

This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.

ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use 0.1µg 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.
  • Target information above from: UniProt accession Q09472 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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 - ChIP Grade images

  • Overlay histogram showing HeLa cells stained with ab54984 (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 (ab54984, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • p300 antibody (ab54984) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human salivary gland.
  • Western blot against tagged recombinant protein immunogen using ab54984 p300 antibody at 1ug/ml. Predicted band size of immunogen is 37 kDa
  • ab54984 at 10 ug/ml staining KAT3B / p300 in human Hela cells by Immunocytochemistry / Immunofluorescence.

References for Anti-KAT3B / p300 antibody - ChIP Grade (ab54984)

This product has been referenced in:
  • Zemans RL  et al. Role of ß-catenin-regulated CCN matricellular proteins in epithelial repair after inflammatory lung injury. Am J Physiol Lung Cell Mol Physiol 304:L415-27 (2013). Read more (PubMed: 23316072) »
  • Li Y  et al. High expression of p300 has an unfavorable impact on survival in resectable esophageal squamous cell carcinoma. Ann Thorac Surg 91:1531-8 (2011). Read more (PubMed: 21524463) »

See all 2 Publications for this product

Product Wall

Application Flow Cytometry
Fixation Formaldehyde
Permeabilization Yes - BD PERMEABILISATION BUFFER
Sample Human Cell (MCF7)
Specification MCF7
Gating Strategy CD44 POSITIVE
Preparation Cell harvesting/tissue preparation method: treated with trypsin to make single cell suspension
Sample buffer: 0.1% PBS BSA
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Submitted Dec 10 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: sodium citrate
Sample Human Tissue sections (BREAST CANCER TISSUE)
Specification BREAST CANCER TISSUE
Permeabilization Yes - xylene
Fixative Formaldehyde
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Submitted Dec 04 2013

Application ChIP
Detection step Semiquantitative PCR
Sample Human Cell lysate - nuclear (MCF7)
Specification MCF7
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: FORMALDEHYDE 10%
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Submitted Dec 04 2013

Application Immunoprecipitation
Immuno-precipitation step Staph. aureus
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Total protein in input 100 µg
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Submitted Sep 06 2013

Application ChIP
Detection step Semiquantitative PCR
Sample Human Cell lysate - nuclear (MCF7)
Specification MCF7
Negative control IgG PROCESSED sample
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: FORMALDEHYDE 10%
Positive control input sample which is processed with out antibody and decrosslinked precipitated DNA
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Submitted Sep 06 2013

Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Human Cell lysate - whole cell (HEK293T)
Specification HEK293T
Total protein in input 100 µg
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Submitted Jul 03 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step 10% NON IMMUNE SERUM as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: sodium citrate
Sample Mouse Tissue sections (MCF7 CELLS INJECTED IN TO MOUSE)
Specification MCF7 CELLS INJECTED IN TO MOUSE
Permeabilization Yes - xylene
Fixative Formaldehyde
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Submitted Jul 03 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jun 24 2013

Abreviews
Application Immunoprecipitation
Immuno-precipitation step Other - sepharose beads
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Total protein in input 100 µg
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Submitted Jun 17 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated
Sample Human Tissue sections (urinary tract)
Specification urinary tract
Permeabilization No
Fixative Formaldehyde
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Submitted May 24 2013

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