Overview

  • Product nameAnti-HDAC3 antibodySee all HDAC3 primary antibodies ...
  • Description
    Rabbit polyclonal to HDAC3
  • Tested applicationsIHC-P, ICC/IF, ICC, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Indian Muntjac, Zebrafish
    Predicted to work with: Chicken, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human HDAC3.

    (Peptide available as ab16279.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A431; HEK293 and Hela Nuclear lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab16047 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
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
ICC 1/100 - 1/500.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).Can be blocked with Human HDAC3 peptide (ab16279).
IP 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
    • 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 images

  • Lanes 1 & 6 : Anti-HDAC3 antibody (ab16047) at 1 µg/ml (Sample: HeLa nuclear lysate, 20 ug)
    Lanes 2 & 7 : Anti-HDAC3 antibody (ab16047) at 1 µg/ml (Sample: HeLa whole cell lysate, 20 ug)
    Lanes 3 & 8 : Anti-HDAC3 antibody (ab16047) at 1 µg/ml (Sample: A431 cell lysate, 20 ug)
    Lanes 4 & 9 : Anti-HDAC3 antibody (ab16047) at 1 µg/ml (Sample: Jurkat cell lysate, 20 ug)
    Lanes 5 & 10 : Anti-HDAC3 antibody (ab16047) at 1 µg/ml (Sample: HEK293 cell lysate, 20 ug)

    Lane 1 : As above
    Lane 2 : As above
    Lane 3 : As above
    Lane 4 : As above
    Lane 5 : As above
    Lane 6 : Human HDAC3 peptide (ab16279) at 1 µg/ml
    Lane 7 : Human HDAC3 peptide (ab16279) at 1 µg/ml
    Lane 8 : Human HDAC3 peptide (ab16279) at 1 µg/ml
    Lane 9 : Human HDAC3 peptide (ab16279) at 1 µg/ml
    Lane 10 : Human HDAC3 peptide (ab16279) at 1 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 50 kDa
    Observed band size : 49 kDa (why is the actual band size different from the predicted?)
  • SK-N-SH cells were fixed in 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab16047 (1/100). The antibody clearly stained the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.

  • Interphase HeLa cells incubated with ab16047 (1/500). The antibody shows predominantly nuclear staining. The ab16047 staining is shown in green. The cells were counterstained with DAPI (red).

     

  • ab16047 staining HDAC3 in human breast cancer tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/2000 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.

    See Abreview

  • All lanes : Anti-HDAC3 antibody (ab16047) at 1 µg/ml

    Lane 1 : Mouse 3T3 cell lysate
    Lane 2 : Rat liver cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa
    Observed band size : 49 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    ab16047 cross-reacts with Mouse 3T3 cell lysate but shows no cross-reactivity against Rat liver lysate.

  • HDAC3 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HDAC3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab16047.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 49kDa: HDAC3.
  • All lanes : Anti-HDAC3 antibody (ab16047) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate (20ug)
    Lane 3 : Zebrafish heart homogenate (20ug)
    Lane 4 : Zebrafish liver homogenate (20ug)

    Secondary
    Donkey polyclonal to Goat IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa
    Observed band size : 48 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

References for Anti-HDAC3 antibody (ab16047)

This product has been referenced in:
  • Takase K  et al. Monoaminergic and neuropeptidergic neurons have distinct expression profiles of histone deacetylases. PLoS One 8:e58473 (2013). ICC/IF ; Mouse . Read more (PubMed: 23469282) »
  • Chu YL  et al. MutS homologue hMSH4: interaction with eIF3f and a role in NHEJ-mediated DSB repair. Mol Cancer 12:51 (2013). WB . Read more (PubMed: 23725059) »

See all 8 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Macrophage)
Specification Macrophage
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Apr 10 2013

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...

Read More
Application Western blot
Sample Xenopus laevis Tissue lysate - whole (tectum)
Loading amount 20 µg
Specification tectum
Gel Running Conditions Reduced Denaturing (MINI-PROTEAN TGX)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. WANHUA SHEN

Verified customer

Submitted Apr 02 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human glioblastoma cell lines)
Specification human glioblastoma cell lines
Fixative Paraformaldehyde
Permeabilization Yes - Tritonx100 0.1% in PBS
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 11 2011

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (breast cancer)
Specification breast cancer
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Jan 22 2009

Application Western blot
Sample Human Cell lysate - whole cell (various cell types - cell lines and human primary)
Loading amount 15 µg
Specification various cell types - cell lines and human primary
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Prof. Paul Townsend

Verified customer

Submitted Apr 21 2008

Application Immunocytochemistry
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Dec 02 2005

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