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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Acetylation
使用敲除细胞株进行验证重组RabMAb

重组Anti-HDAC2抗体[Y461] (ab32117)

  • Datasheet
Reviews (11)Q&A (8)References (50)

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Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] (ab32117)
  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] (ab32117)
  • Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
  • Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
  • Immunoprecipitation - Anti-HDAC2 antibody [Y461] (ab32117)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
  • Flow Cytometry - Anti-HDAC2 antibody [Y461] (ab32117)
  • Anti-HDAC2 antibody [Y461] (ab32117)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y461] to HDAC2
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

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概述

  • 产品名称

    Anti-HDAC2抗体[Y461]
    参阅全部 HDAC2 一抗
  • 描述

    兔单克隆抗体[Y461] to HDAC2
  • 宿主

    Rabbit
  • 特异性

    ab32117 recognises HDAC2.
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • 免疫原

    Synthetic peptide within Human HDAC2 aa 450-550 (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: HAP1, A431, Hela and K562 cell lysate and rat brain tissue homogenate. IHC-P: Human breast carcinoma and rat spinal cord tissue. ICC/IF: MCF-7 and wildtype HAP1 cells. Flow Cyt: HeLa cells.
  • 常规说明

    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.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 克隆

    单克隆
  • 克隆编号

    Y461
  • 同种型

    IgG
  • 研究领域

    • 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
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other

相关产品

  • Alternative Versions

    • HRP Anti-HDAC2 antibody [Y461] (ab195851)
    • Alexa Fluor® 488 Anti-HDAC2 antibody [Y461] (ab196471)
    • Alexa Fluor® 647 Anti-HDAC2 antibody [Y461] (ab196518)
    • PE Anti-HDAC2 antibody [Y461] (ab210827)
    • Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    • Alexa Fluor® 568 Anti-HDAC2 antibody [Y461] (ab214384)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

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

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

    • Recombinant human HDAC2 protein (ab42630)

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32117 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Human
All applications
Mouse
Rat
应用 Ab评论 说明
WB (9)
1/2000. Predicted molecular weight: 55 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/250 - 1/500.
Flow Cyt
1/60 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP
1/60.
说明
WB
1/2000. Predicted molecular weight: 55 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/250 - 1/500.
Flow Cyt
1/60 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP
1/60.

靶标

  • 功能

    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.
    Forms transcriptional repressor complexes by associating with MAD, SIN3, YY1 and N-COR. Interacts in the late S-phase of DNA-replication with DNMT1 in the other transcriptional repressor complex composed of DNMT1, DMAP1, PCNA, CAF1. Deacetylates TSHZ3 and regulates its transcriptional repressor activity.
  • 组织特异性

    Widely expressed; lower levels in brain and lung.
  • 序列相似性

    Belongs to the histone deacetylase family. HD type 1 subfamily.
  • 翻译后修饰

    S-nitrosylated by GAPDH. In neurons, S-Nitrosylation at Cys-262 and Cys-274 does not affect the enzyme activity but abolishes chromatin-binding, leading to increases acetylation of histones and activate genes that are associated with neuronal development. In embryonic cortical neurons, S-Nitrosylation regulates dendritic growth and branching.
  • 细胞定位

    Nucleus.
  • Target information above from: UniProt accession Q92769 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 3066 Human
    • Entrez Gene: 15182 Mouse
    • Entrez Gene: 84577 Rat
    • Omim: 605164 Human
    • SwissProt: Q92769 Human
    • SwissProt: P70288 Mouse
    • Unigene: 3352 Human
    • Unigene: 19806 Mouse
    • Unigene: 1797 Rat
    see all
  • 别名

    • D10Wsu179e antibody
    • HD 2 antibody
    • HD2 antibody
    • HDAC 2 antibody
    • Hdac2 antibody
    • HDAC2_HUMAN antibody
    • Histone deacetylase 2 (HD2) antibody
    • Histone deacetylase 2 antibody
    • OTTHUMP00000017046 antibody
    • OTTHUMP00000227077 antibody
    • OTTHUMP00000227078 antibody
    • RPD3 antibody
    • transcriptional regulator homolog RPD3 antibody
    • YAF1 antibody
    • YY1 associated factor 1 antibody
    • YY1 transcription factor binding protein antibody
    • Yy1bp antibody
    see all

图片

  • Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
    Western blot - Anti-HDAC2 antibody [Y461] (ab32117)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: HDAC2 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: A431 whole cell lysate (20 µg)
    Lane 4: Hela whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32117 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32117 was shown to specifically react with HDAC2 when HDAC2 knockout samples were used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE.  ab32117 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 and 1/10000 respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] (ab32117)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] (ab32117)

    ab32117 staining HDAC2 in wild-type HAP1 cells (top panel) and HDAC2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32117 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] (ab32117)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] (ab32117)

    ab32117 staining HDAC2 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

    Negative control 1: PBS only.

  • Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
    Western blot - Anti-HDAC2 antibody [Y461] (ab32117)This image is courtesy of an anonymous Abreview
    All lanes : Anti-HDAC2 antibody [Y461] (ab32117) at 1/1000 dilution

    Lane 1 : Rat brain tissue homogenate at 20 µg
    Lane 2 : Rat brain tissue homogenate at 40 µg
    Lane 3 : Rat brain tissue homogenate, P1 nuclear fraction at 20 µg
    Lane 4 : Rat brain tissue homogenate, P1 nuclear fraction at 40 µg
    Lane 5 : Rat brain tissue homogenate, P2 non-nuclear fraction at 20 µg
    Lane 6 : Rat brain tissue homogenate, P2 non-nuclear fraction at 40 µg

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/1000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 35 kDa (possible non-specific binding)


    Exposure time: 5 minutes

    See Abreview

  • Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
    Western blot - Anti-HDAC2 antibody [Y461] (ab32117)
    Anti-HDAC2 antibody [Y461] (ab32117) at 1/2000 dilution + K562 cell lysate

    Predicted band size: 55 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?

  • Immunoprecipitation - Anti-HDAC2 antibody [Y461] (ab32117)
    Immunoprecipitation - Anti-HDAC2 antibody [Y461] (ab32117)

    HDAC2 was immunoprecipitated from 1mg of Hela(Human epithelial cell line from cervix adenocarcinoma)whole cell lysate with ab32117at 1/50 dilution.

    Western blot was performed from the immunoprecipitate using ab32117 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP),specific to the non-reduced form of IgG, was used as secondary antibody at 1/1000 dilution.

    Lane 1: Hela whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)

    Immunohistochemical analysis of HDAC2 expression in paraffin embedded human breast carcinoma tissue section, using 1/250 ab32117.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-HDAC2 antibody [Y461] (ab32117)
    Flow Cytometry - Anti-HDAC2 antibody [Y461] (ab32117)
    Overlay histogram showing HeLa cells stained with ab32117 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32117, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Anti-HDAC2 antibody [Y461] (ab32117)
    Anti-HDAC2 antibody [Y461] (ab32117)

实验方案

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

数据表及文件

    • Datasheet
  • 文献 (50)

    发表研究结果有使用 ab32117?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab32117 被引用在 50 文献中.

    • Lin YH  et al. HDAC2 (Histone deacetylase 2): A critical factor in environmental enrichment-mediated stroke recovery. J Neurochem N/A:N/A (2020). PubMed: 32415988
    • Basta J  et al. Pharmacologic inhibition of RGD-binding integrins ameliorates fibrosis and improves function following kidney injury. Physiol Rep 8:e14329 (2020). PubMed: 32281744
    • Meng Q  et al. Protective effects of histone deacetylase inhibition by Scriptaid on brain injury in neonatal rat models of cerebral ischemia and hypoxia. Int J Clin Exp Pathol 13:179-191 (2020). PubMed: 32211098
    • Chung J  et al. Protein arginine methyltransferase 5 (PRMT5) promotes survival of lymphoma cells via activation of WNT/ß-catenin and AKT/GSK3ß proliferative signaling. J Biol Chem 294:7692-7710 (2019). PubMed: 30885941
    • Oka OB  et al. ERp18 regulates activation of ATF6a during unfolded protein response. EMBO J 38:e100990 (2019). PubMed: 31368601
    View all Publications for this product

    客户评价及客户问答

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    1-10 of 11 Abreviews

    Immunohistochemistry (Frozen sections) abreview for Anti-HDAC2 antibody [Y461]

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    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (HF)
    Permeabilization
    Yes - PBS-TWEEN 0.05%
    Specification
    HF
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

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    Verified customer

    提交于 Feb 28 2018

    Western blot abreview for Anti-HDAC2 antibody [Y461]

    Excellent
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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (esophagus)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    40 µg
    Specification
    esophagus
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 05 2016

    Western blot abreview for Anti-HDAC2 antibody [Y461]

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    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (lung homogenates)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Specification
    lung homogenates
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 25 2015

    Western blot abreview for Anti-HDAC2 antibody [Y461]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Peridontal ligament fibroblast)
    Gel Running Conditions
    Reduced Denaturing (7.5%)
    Loading amount
    20 µg
    Specification
    Peridontal ligament fibroblast
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 15 2015

    Western blot abreview for Anti-HDAC2 antibody [Y461]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Sample
    Rat Tissue lysate - nuclear (Brain tissue homogenate, P1 and P2 fractions)
    Specification
    Brain tissue homogenate, P1 and P2 fractions
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 31 2014

    Immunohistochemistry (Frozen sections) abreview for Anti-HDAC2 antibody [Y461]

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    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Spinal cord)
    Specification
    Spinal cord
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 24 2010

    Western blot abreview for Anti-HDAC2 antibody [Y461]

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Loading amount
    100 µg
    Specification
    HeLa
    Treatment
    200ng/ml Nocodazole for 12hr
    Gel Running Conditions
    Reduced Denaturing (8% acrylamide gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More

    Abcam user community

    Verified customer

    提交于 Nov 20 2009

    Western blot abreview for Anti-HDAC2 antibody [Y461]

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (HeLa cells)
    Loading amount
    10 µg
    Specification
    HeLa cells
    Gel Running Conditions
    Reduced Denaturing (7)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 08 2008

    Western blot abreview for Anti-HDAC2 antibody [Y461]

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human aortic smooth muscle cells)
    Loading amount
    5 µg
    Specification
    human aortic smooth muscle cells
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Dec 22 2007

    Western blot abreview for Anti-HDAC2 antibody [Y461]

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - nuclear (Lung)
    Loading amount
    10 µg
    Specification
    Lung
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    提交于 Aug 23 2007

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