Overview

  • Product nameAnti-HDAC3 antibody - ChIP GradeSee 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:

    C-NEFYDGDHDNDKESDVEI

    conjugated to KLH, corresponding to amino acids 411-428 of Human HDAC3.

  • EpitopeThe epitope recognized by the antibody is resistant to routine formalin-fixation and paraffin-embedding.

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 dilution.
ChIP Use at an assay dependent dilution. Use 2µl for 2 x 106 cells.
ICC/IF Use at an assay dependent dilution.

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
    • HD 3 antibody
    • HD3 antibody
    • HDAC 3 antibody
    • HDAC3 antibody
    • HDAC3_HUMAN antibody
    • Histone deacetylase 3 (HD3) (RPD3-2) antibody
    • histone deacetylase 3 antibody
    • RPD 3 antibody
    • RPD3 2 antibody
    • RPD3 antibody
    • RPD3-2 antibody
    • SMAP 45 antibody
    • SMAP45 antibody
    see all

Anti-HDAC3 antibody - ChIP Grade images

  • 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 2x106 cells were used in each ChIP experiment.

  • 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 TBS-T. Coverslips were then counterstained with DAPI in TBS-T for 1-2 minutes, TBS-T was then added and the coverslips mounted. Red indicates staining by ab7030, blue by DAPI. A small amount of cytoplasmic staining was seen with ab7030.
  • 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:
  • 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) »
  • Shen W  et al. Epigenetic modification of the leptin promoter in diet-induced obese mice and the effects of N-3 polyunsaturated Fatty acids. Sci Rep 4:5282 (2014). ChIP ; Mouse . Read more (PubMed: 24923522) »

See all 23 Publications for this product

Product Wall

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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 (...

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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

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

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

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

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

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

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Breast Tumour)
Specification Breast Tumour
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated
Blocking step Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 20
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Submitted Jul 27 2006

Application Western blot
Sample Human Cell lysate - whole cell (Liver Cell Line - LX2)
Loading amount 30 µg
Specification Liver Cell Line - LX2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
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Dr. Ahmed Elsharkawy

Verified customer

Submitted Jul 14 2006

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