Recombinant
RabMAb

Anti-HDAC3 antibody [Y415] (ab32369)

Overview

  • Product name
    Anti-HDAC3 antibody [Y415]
    See all HDAC3 primary antibodies
  • Description
    Rabbit monoclonal [Y415] to HDAC3
  • Specificity
    This antibody may detect splice isoform 2 (RPD3-2A).
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human HDAC3 (C terminal).

  • Positive control
    • K562 cell lysate; HeLa cells; human ovary carcinoma
  • General notes

    This product is a recombinant rabbit monoclonal antibody. Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab32369 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
Flow Cyt 1/30.
WB 1/5000 - 1/10000. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).Can be blocked with HDAC3 peptide (ab221015).
IHC-P 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified use at 1/250 - 1/500. 

 See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol

ICC/IF 1/250 - 1/500.
IP 1/20 - 1/50.

Target

  • Function
    Responsible 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 specificity
    Widely expressed.
  • Sequence similarities
    Belongs to the histone deacetylase family. HD type 1 subfamily.
  • Post-translational
    modifications
    Sumoylated in vitro.
  • Cellular localization
    Nucleus.
  • 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 [Y415] images

  • All lanes : Anti-HDAC3 antibody [Y415] (ab32369) at 1/5000 dilution

    Lane 1 : K562 (human chronic myelogenous leukemia) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryo) whole cell lysate
    Lane 4 : C6 (rat glioma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size : 49 kDa
    Observed band size : 49 kDa
    Blocking/Diluting buffer 5% NFDM/TBST
  • Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma sections labelling HDAC3 with purified ab32369 at dilution of 1:50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence analysis of K562 cells labelling HDAC3 with purified ab32369 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. ab150077, Alexa Fluor®488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor®594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei were counterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used and for negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used. 

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling HDAC3 with purified ab32369 at 1/30 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Immunofluorescent staining of HeLa cells
    using ab32369 at 1/250 dilution
  • Purified ab32369 at 1/20 immunoprecipitating HDAC3 in K562 whole cell lysate observed at 49 KDa (lanes 1 and 2). 

    Lane 1 (input): K562 whole cell lysate 10ug              

    Lane 2 (+): ab32369 + K562 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32369 in K562 whole cell lysate 

    The secondary antibody used was VeriBlot for IP (HRP) (ab131366) at dilution of 1/1000.            

    Blocking/Diluting buffer 5% NFDM/TBST. 

  • Anti-HDAC3 antibody [Y415] (ab32369) at 1/10000 dilution + K562 cell lysate

    Predicted band size : 49 kDa
    Observed band size : 49 kDa
  • Paraffin-embedded human ovary carcinoma
    ab32369 at 1/250 dilutionParaffin-embedded human ovary carcinoma
    ab32369 at 1/250 dilution

References for Anti-HDAC3 antibody [Y415] (ab32369)

This product has been referenced in:
  • Huang R  et al. The role of HDAC2 in chromatin remodelling and response to chemotherapy in ovarian cancer. Oncotarget 7:4695-711 (2016). WB, IHC-P ; Human . Read more (PubMed: 26683361) »
  • Jamaladdin S  et al. Histone deacetylase (HDAC) 1 and 2 are essential for accurate cell division and the pluripotency of embryonic stem cells. Proc Natl Acad Sci U S A 111:9840-5 (2014). Mouse . Read more (PubMed: 24958871) »

See all 7 Publications for this product

Product Wall

Application
Western blot
Sample
Mouse Tissue lysate - whole (lung)
Gel Running Conditions
Reduced Denaturing (7.5)
Loading amount
30 µg
Treatment
Oxidant 24hr
Specification
lung
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Dec 08 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (Brain)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
12 µg
Specification
Brain
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jun 01 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (embryo)
Specification
embryo
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate based, high pH (Vector lab)
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 02 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lymph node)
Specification
Lymph node
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer, pH6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 01 2011

Thank you for your enquiry. I understand your concern. There does, however, appear to be some evidence that HDAC3 can be found in the cytosol. Please see Figure 6 in the paper found at the following link: http://www.pubmedcentral.nih.gov/articl...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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 minute(s) · Concentration: 20
Username

Abcam user community

Verified customer

Submitted Jul 28 2006

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

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