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  1. Link

    hdac2-antibody-epr5001-chip-grade-ab124974.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Acetylation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)

  • Datasheet
  • SDS
Reviews (4) Submit a question References (9)

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Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • ChIP - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
  • Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5001] to HDAC2 - ChIP Grade
  • Suitable for: WB, ICC/IF, ChIP
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Knockout
Product image
Human HDAC2 knockout HEK-293T cell lysate (ab256939)
Knockout
Product image
Human HDAC2 knockout HEK-293T cell line (ab266588)
Knockout
Product image
Human HDAC2 knockout HEK-293T cell line (ab266589)

View more associated products

Overview

  • Product name

    Anti-HDAC2 antibody [EPR5001] - ChIP Grade
    See all HDAC2 primary antibodies
  • Description

    Rabbit monoclonal [EPR5001] to HDAC2 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, ChIPmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HEK-293T, HAP1, HeLa, A431, SH-SY5Y, and Jurkat cell lysates. ICC/IF: Wild-type HAP1 cells. ChIP: Chromatin extract from HeLa cells.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5001
  • 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
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Anti-HDAC2 antibody [EPR5001] - BSA and Azide free (ab248081)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human HDAC2 knockout HEK-293T cell line (ab266588)
    • Human HDAC2 knockout HEK-293T cell line (ab266589)
    • Human HDAC2 knockout HEK-293T cell line (ab266590)
  • KO cell lysates

    • Human HDAC2 knockout HEK-293T cell lysate (ab256937)
    • Human HDAC2 knockout HEK-293T cell lysate (ab256938)
    • Human HDAC2 knockout HEK-293T cell lysate (ab256939)
  • Positive Controls

    • A-431 whole cell lysate (ab30132)
    • Jurkat whole cell lysate (ab7899)
    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant human HDAC2 protein (ab42630)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124974 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
WB (3)
1/10000 - 1/50000. Predicted molecular weight: 55 kDa.
ICC/IF
Use a concentration of 0.5 µg/ml.
ChIP (1)
Use 10 µg for 25 µg of chromatin.

Please note product formulation is not optimised for ChIP application

Notes
WB
1/10000 - 1/50000. Predicted molecular weight: 55 kDa.
ICC/IF
Use a concentration of 0.5 µg/ml.
ChIP
Use 10 µg for 25 µg of chromatin.

Please note product formulation is not optimised for ChIP application

Application notes
Is unsuitable for Flow Cyt or IHC-P.

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.
    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.
  • Tissue specificity

    Widely expressed; lower levels in brain and lung.
  • Sequence similarities

    Belongs to the histone deacetylase family. HD type 1 subfamily.
  • Post-translational
    modifications

    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.
  • Cellular localization

    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
  • Database links

    • 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
    • Alternative names

      • 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

    Images

    • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : HDAC2 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 55 kDa
      Observed band size: 55 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab124974 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab124974 was shown to react with HDAC2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266589 (knockout cell lysate ab256938) was used. Wild-type HEK-293T and HDAC2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)

      ab124974 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 ab124974 at 0.5μg/ml 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).

    • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : HDAC2 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 55 kDa
      Observed band size: 60 kDa why is the actual band size different from the predicted?



      Lanes 1-2: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : HDAC2 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 55 kDa
      Observed band size: 60 kDa why is the actual band size different from the predicted?



      Lanes 1-2: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266588 (knockout cell lysate ab256937) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : HDAC2 knockout HAP1 whole cell lysate
      Lane 3 : A431 whole cell lysate
      Lane 4 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 55 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab124974 was shown to recognize HDAC2 when HDAC2 knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE.  ab124974 and ab8245 (Mouse anti GAPDH loading control) were but diluted at 1/10000 and incubated overnight at 4°C. 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.

       

    • Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : SH-SY5Y cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : A431 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 55 kDa

    • ChIP - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      ChIP - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)

      Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 10µg of ab124974 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.

    • Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
      Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)

    Protocols

    • ChIP protocols
    • Western blot protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (9)

    Publishing research using ab124974? Please let us know so that we can cite the reference in this datasheet.

    ab124974 has been referenced in 9 publications.

    • Chen Z  et al. Imaging assisted evaluation of antitumor efficacy of a new histone deacetylase inhibitor in the castration-resistant prostate cancer. Eur J Nucl Med Mol Imaging 48:53-66 (2021). PubMed: 32592040
    • Xie L  et al. Elucidation of the Hdac2/Sp1/miR-204-5p/Bcl-2 axis as a modulator of cochlear apoptosis via in vivo/in vitro models of acute hearing loss. Mol Ther Nucleic Acids 23:1093-1109 (2021). PubMed: 33614251
    • Pan L  et al. Silencing of CREB Inhibits HDAC2/TLR4/NF-?B Cascade to Relieve Severe Acute Pancreatitis-Induced Myocardial Injury. Inflammation 44:1565-1580 (2021). PubMed: 33725236
    • Xu L  et al. H3K14 hyperacetylation-mediated c-Myc binding to the miR-30a-5p gene promoter under hypoxia postconditioning protects senescent cardiomyocytes from hypoxia/reoxygenation injury. Mol Med Rep 23:N/A (2021). PubMed: 33880587
    • Gilbert TM  et al. Neuroepigenetic signatures of age and sex in the living human brain. Nat Commun 10:2945 (2019). PubMed: 31270332
    • Link S  et al. PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex. Nat Commun 9:4300 (2018). PubMed: 30327463
    • Hanigan TW  et al. Scaffold dependent histone deacetylase (HDAC) inhibitor induced re-equilibration of the subcellular localization and post-translational modification state of class I HDACs. PLoS One 12:e0186620 (2017). PubMed: 29045501
    • Hanigan TW  et al. Divergent JNK Phosphorylation of HDAC3 in Triple-Negative Breast Cancer Cells Determines HDAC Inhibitor Binding and Selectivity. Cell Chem Biol 24:1356-1367.e8 (2017). PubMed: 28943357
    • Lin MY  et al. Redirection of Epithelial Immune Responses by Short-Chain Fatty Acids through Inhibition of Histone Deacetylases. Front Immunol 6:554 (2015). WB ; Human . PubMed: 26579129

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    ChIP abreview for Anti-HDAC2 antibody [EPR5001] - ChIP Grade

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (K562)
    Specification
    K562
    Detection step
    Other
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% Formaldehyde
    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.

    MR. Andrew Perez

    Verified customer

    Submitted Mar 24 2022

    Western blot abreview for Anti-HDAC2 antibody [EPR5001] - ChIP Grade

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (10% Gel)
    Loading amount
    20 µg
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · 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 Mar 28 2017

    Western blot abreview for Anti-HDAC2 antibody [EPR5001] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    12 µg
    Specification
    Brain
    Blocking step
    BSA 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 25 2015

    Western blot abreview for Anti-HDAC2 antibody [EPR5001]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Monkey 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 May 29 2015

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