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  1. Link

    hdac3-antibody-y415-ab32369.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Acetylation
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RecombinantRabMAb

Recombinant Anti-HDAC3 antibody [Y415] (ab32369)

  • Datasheet
  • SDS
Reviews (6)Q&A (1)References (41)

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Western blot - Anti-HDAC3 antibody [Y415] (ab32369)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)
  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)
  • Flow Cytometry (Intracellular) - Anti-HDAC3 antibody [Y415] (ab32369)
  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)
  • Immunoprecipitation - Anti-HDAC3 antibody [Y415] (ab32369)
  • Western blot - Anti-HDAC3 antibody [Y415] (ab32369)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)
  • Anti-HDAC3 antibody [Y415] (ab32369)

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 (Intra), WB, IHC-P, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free HRP

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Overview

  • Product name

    Anti-HDAC3 antibody [Y415]
    See all HDAC3 primary antibodies
  • Description

    Rabbit monoclonal [Y415] to HDAC3
  • Host species

    Rabbit
  • Specificity

    This antibody may detect splice isoform 2 (RPD3-2A).
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human HDAC3 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab221015)

  • Positive control

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

    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
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.12% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y415
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • HDACs
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • HDACs
    • Class I
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • HRP Anti-HDAC3 antibody [Y415] (ab208356)
    • Anti-HDAC3 antibody [Y415] - BSA and Azide free (ab219376)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant human HDAC3 protein (ab101663)

Applications

The Abpromise guarantee

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 (Intra)
1/30.
WB (3)
1/5000 - 1/10000. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).
IHC-P (3)
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 IHC antigen retrieval protocols.

ICC/IF
1/250 - 1/500.
IP
1/20 - 1/50.
Notes
Flow Cyt (Intra)
1/30.
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. 

See IHC antigen retrieval protocols.

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.
  • Target information above from: UniProt accession O15379 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8841 Human
    • Entrez Gene: 15183 Mouse
    • Entrez Gene: 84578 Rat
    • Omim: 605166 Human
    • SwissProt: O15379 Human
    • SwissProt: O88895 Mouse
    • SwissProt: Q6P6W3 Rat
    • Unigene: 519632 Human
    • Unigene: 20521 Mouse
    • Unigene: 17284 Rat
    see all
  • 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

Images

  • Western blot - Anti-HDAC3 antibody [Y415] (ab32369)
    Western blot - Anti-HDAC3 antibody [Y415] (ab32369)
    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
    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
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)

    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 - Anti-HDAC3 antibody [Y415] (ab32369)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)

    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 (Intracellular) - Anti-HDAC3 antibody [Y415] (ab32369)
    Flow Cytometry (Intracellular) - Anti-HDAC3 antibody [Y415] (ab32369)

    Intracellular 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 Fluorr®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. 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)
    Immunofluorescent staining of HeLa cells
    using ab32369 at 1/250 dilution
  • Immunoprecipitation - Anti-HDAC3 antibody [Y415] (ab32369)
    Immunoprecipitation - Anti-HDAC3 antibody [Y415] (ab32369)

    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. 

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

    Predicted band size: 49 kDa
    Observed band size: 49 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)

    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.

  • Anti-HDAC3 antibody [Y415] (ab32369)
    Anti-HDAC3 antibody [Y415] (ab32369)

Protocols

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.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (41)

Publishing research using ab32369? Please let us know so that we can cite the reference in this datasheet.

ab32369 has been referenced in 41 publications.

  • Na J  et al. The crosstalk of HDAC3, microRNA-18a and ADRB3 in the progression of heart failure. Cell Biosci 11:31 (2021). PubMed: 33549119
  • Huang JW  et al. MicroRNA-328-5p Alleviates Macrophage Lipid Accumulation through the Histone Deacetylase 3/ATP-binding cassette transporter A1 pathway. Lipids 56:301-311 (2021). PubMed: 33663010
  • Hu Q  et al. Histone Deacetylase 3 Aggravates Type 1 Diabetes Mellitus by Inhibiting Lymphocyte Apoptosis Through the microRNA-296-5p/Bcl-xl Axis. Front Genet 11:536854 (2020). PubMed: 33240312
  • Yu XH  et al. LncRNA kcnq1ot1 promotes lipid accumulation and accelerates atherosclerosis via functioning as a ceRNA through the miR-452-3p/HDAC3/ABCA1 axis. Cell Death Dis 11:1043 (2020). PubMed: 33293505
  • Tao H  et al. Coordinated expression of p300 and HDAC3 upregulates histone acetylation during dentinogenesis. J Cell Biochem 121:2478-2488 (2020). PubMed: 31692090
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-7 of 7 Abreviews or Q&A

Western blot abreview for Anti-HDAC3 antibody [Y415]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - nuclear (HeLa cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
HeLa cells
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 27 2017

Western blot abreview for Anti-HDAC3 antibody [Y415]

Excellent
Abreviews
Abreviews
abreview image
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%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 08 2015

Western blot abreview for Anti-HDAC3 antibody [Y415]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 01 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HDAC3 antibody [Y415]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 02 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HDAC3 antibody [Y415]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 01 2011

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HDAC3 antibody [Y415]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 28 2006

Question

I am seeing cytoplasmic staining as well as nuclear staining in ICC, and a signal at the expected MW in Western blots of cytosolic cell fractions. Blots for lamin confirm that the fractionation procedure was correct; there is no lamin detected in the cytosolic fraction.

Read More

Abcam community

Verified customer

Asked on Sep 28 2007

Answer

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/articlerender.fcgi?artid=1618574 Am J Pathol. 2004 August; 165(2): 553–564. There may be other papers referenced in this article that discuss the same observation. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Sep 28 2007

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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