Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)

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 - BSA and Azide free
  • Suitable for: WB, IP, ICC/IF, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-HDAC2 antibody [Y461] - BSA and Azide free
    See all HDAC2 primary antibodies

  • Description

    Rabbit monoclonal [Y461] to HDAC2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • Human breast carcinoma, K562 cells, HeLa cells.

  • General notes

    Ab213700 is the carrier-free version of ab32117. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab213700 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Western blot - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    All lanes : HRP Anti-HDAC2 antibody [Y461] (ab195851) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : HDAC2 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 55 kDa



    ab195851 was shown to specifically react with HDAC2 in wild-type HAP1 cells as signal was lost in HDAC2 knockout cells. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. Ab195851 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195851).

  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32117).

  • Immunoprecipitation - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Immunoprecipitation - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32117).

  • Flow Cytometry - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Flow Cytometry - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32117).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32117).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)

    This ICC data was generated using the same anti-HDAC2 antibody clone, Y461, in a different buffer formulation (cat# 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).

     

  • Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
    Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)

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