Overview

  • Product nameAnti-HDAC3 antibody - ChIP Grade
    See all HDAC3 primary antibodies
  • Description
    Rabbit polyclonal to HDAC3 - ChIP Grade
  • SpecificityAnti-Histone Deacetylase 3 specifically recognizes histone deacetylase 3 by immunoblotting and immunoprecipitation (50 kD).
  • Tested applicationsWB, IP, IHC-P, Dot Blot, ChIP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human HDAC3 aa 411-428 conjugated to Keyhole Limpet Haemocyanin (KLH). The epitope recognized by the antibody is resistant to routine formalin-fixation and paraffin-embedding.
    Sequence:

    C-NEFYDGDHDNDKESDVEI

Properties

Applications

Our Abpromise guarantee covers the use of ab7030 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).
IP 1/2500.
IHC-P 1/500.
Dot Blot Use at an assay dependent concentration.
ChIP Use 2µl for 106 cells. PubMed: 18830415
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionResponsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Probably participates in the regulation of transcription through its binding to the zinc-finger transcription factor YY1; increases YY1 repression activity. Required to repress transcription of the POU1F1 transcription factor. Acts as a molecular chaperone for shuttling phosphorylated NR2C1 to PML bodies for sumoylation.
  • Tissue specificityWidely expressed.
  • Sequence similaritiesBelongs to the histone deacetylase family. HD type 1 subfamily.
  • Post-translational
    modifications
    Sumoylated in vitro.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • HD3 antibody
    • HDAC 3 antibody
    • HDAC3 antibody
    • HDAC3_HUMAN antibody
    • Histone deacetylase 3 antibody
    • RPD3 2 antibody
    • RPD3 antibody
    • RPD3-2 antibody
    • SMAP45 antibody
    see all

Anti-HDAC3 antibody - ChIP Grade images

  • All lanes : Anti-HDAC3 antibody - ChIP Grade (ab7030) at 1/5000 dilution

    Lane 1 : HEK-293T whole cell lysate.
    Lane 2 : HeLa whole cell lysate.
    Lane 3 : K562 whole cell lysate.
    Lane 4 : COS-7 whole cell lysate.
    Lane 5 : CHO whole cell lysate.
    Lane 6 : NIH-3T3 whole cell lysate.
    Lane 7 : Neuro-2a whole cell lysate.
    Lane 8 : PC-12 whole cell lysate.
    Lane 9 : NRK whole cell lysate.

    Secondary
    Goat Anti-Rabbit IgG-Peroxidase and a chemiluminescent substrate.

    Predicted band size : 49 kDa
  • Immunocytochemical immunofluorescence analysis of HeLa cells labelling HDAC3 with ab7030. Cells were fixed and permeabilized with cold methanol followed by a cold methanol and acetone solution. Fixed cells were stained with ab7030 at a 1/200 concentration. The secondary used was a Goat Anti-Rabbit IgG, Cy3 conjugate.

  • Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab7030 to HDAC3 and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 2 µl of ab7030 and 2x10

    Cells were fixed with 4% formaldehyde in PEM buffer. The coverslip was incubated in blocking buffer of 5% powdered milk in TBS-T plus 0.02% sodium azide for 1 hour at room temperature. Blocking buffer was removed and primary antibody was added at a dilution of 1/5000 and incubated overnight at 4 degrees celsius. The coverslips were then washed 4-5 times with blocking buffer for 5 minutes. Secondary antibody, goat anti-rabbit Alexa 594, was added at a dilution of 1/1000 and incubated at room temperature for one hour. From this point on coverslips were covered with foil to protect them from light. They were washed 5 times with TBS-T and then one time with PEM, for 5 minutes each wash. The coverslips were fixed 10-30 minutes in 4% formaldehyde in PEM buffer, then washed 3 times with PEM buffer for 5 minutes. 0.1M ammonium chloride in PEM buffer was added for 10 minutes to quench auto-florescence, and then slips were washed 2 times for 5 minutes in PEM followed by 3 washes for 5 minutes in
  • All lanes : Anti-HDAC3 antibody - ChIP Grade (ab7030) at 1/10000 dilution

    Lane 1 : Whole cell lysate from human HEK293 cell line
    Lane 2 : Whole cell lysate from human HEK293 cell line treated with HDAC3 gene silencing shRNA

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit Ig at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 49 kDa


    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-HDAC3 antibody - ChIP Grade (ab7030)

This product has been referenced in:
  • Mani SK  et al. Inhibition of class I histone deacetylase activity represses matrix metalloproteinase-2 and -9 expression and preserves LV function postmyocardial infarction. Am J Physiol Heart Circ Physiol 308:H1391-401 (2015). Read more (PubMed: 25795711) »
  • Kim DW  et al. A histone deacetylase inhibitor improves hypothyroidism caused by a TRa1 mutant. Hum Mol Genet 23:2651-64 (2014). Mouse . Read more (PubMed: 24381310) »

See all 28 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Monkey Cell lysate - whole cell (Brain)
Gel Running Conditions Reduced Denaturing (4-20%)
Loading amount 15 µg
Specification Brain
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted May 29 2015

Application ChIP
Detection step Real-time PCR
Sample Mouse Cell lysate - whole cell (Embryonic Stem Cell)
Specification Embryonic Stem Cell
Negative control Immuno-precipitation with non-specific IgG antibody.
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 60 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Positive control WT embryonic stem cells treated with all-trans retinoic acid (1uM)
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Submitted Feb 04 2015

Thank you for contacting us.

While the specific immunogen for both ab16047 and ab7030 are similar, some antibodies work better in certain applications due to the nature in which they are derived, stored, and their specific affinity for the ant...

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The mouse HDAC3 has 2 isoforms: 27 and 48 kDa (http://www.uniprot.org/uniprot/O88895).
Also, 2 customers had submitted Abreviews for mouse and WB, one of them for 3T3 cells where they also obtained 2 bands (...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (lymphoblastoid cell line)
Specification lymphoblastoid cell line
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step Real-time PCR
Positive control NFKBIA promoter
Negative control GAPDH coding sequence
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Submitted Jul 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (3T3)
Loading amount 500000 cells
Specification 3T3
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Jan 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Mouse brain)
Loading amount 20 µg
Specification Mouse brain
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (lymphoblastoid cell line)
Loading amount 20 µg
Specification lymphoblastoid cell line
Gel Running Conditions Non-reduced Denaturing
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 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (Jurkat Cells)
Specification Jurkat Cells
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
Detection step Real-time PCR
Positive control Total Histone 3 antibody from Upstate
Negative control IgG
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Submitted May 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 20 µg
Specification HEK293
Treatment HDAC3 - shRNA 48 hours
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Oct 10 2008

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