Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y415] to HDAC3
- Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Product nameAnti-HDAC3 antibody [Y415]
See all HDAC3 primary antibodies
DescriptionRabbit monoclonal [Y415] to HDAC3
SpecificityThis antibody may detect splice isoform 2 (RPD3-2A).
Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ICC/IF, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human HDAC3 (C terminal). The exact sequence is proprietary.
(Peptide available as
- K562 cell lysate; HeLa cells; human ovary carcinoma
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.12% BSA
Concentration information loading...
PurityProtein A purified
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
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.
|WB||1/5000 - 1/10000. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).|
|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.
|ICC/IF||1/250 - 1/500.|
|IP||1/20 - 1/50.|
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.
modificationsSumoylated in vitro.
- Information by UniProt
- HD3 antibody
- HDAC 3 antibody
- HDAC3 antibody
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.
All lanes : 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 Detection Reagent (HRP) (ab131366) for detection 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 dilution
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32369 has been referenced in 32 publications.
- Yang C et al. FBP1 binds to the bromodomain of BRD4 to inhibit pancreatic cancer progression. Am J Cancer Res 10:523-535 (2020). PubMed: 32195024
- Cao L et al. Genotoxic stress-triggered ß-catenin/JDP2/PRMT5 complex facilitates reestablishing glutathione homeostasis. Nat Commun 10:3761 (2019). PubMed: 31434880
- Du Y et al. Ubiquitin specific peptidase 5 promotes ovarian cancer cell proliferation through deubiquitinating HDAC2. Aging (Albany NY) 11:9778-9793 (2019). PubMed: 31727867
- Gilbert TM et al. Neuroepigenetic signatures of age and sex in the living human brain. Nat Commun 10:2945 (2019). PubMed: 31270332
- Zhao Q et al. HDAC3 inhibition prevents blood-brain barrier permeability through Nrf2 activation in type 2 diabetes male mice. J Neuroinflammation 16:103 (2019). PubMed: 31101061