Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1698(2)] to HDAC6
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Human, Monkey

Overview

  • Product name

    Anti-HDAC6 antibody [EPR1698(2)]
    See all HDAC6 primary antibodies

  • Description

    Rabbit monoclonal [EPR1698(2)] to HDAC6

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Human, Monkey

  • Immunogen

    Synthetic peptide within Human HDAC6 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q9UBN7

  • Positive control

    • WB: HAP1, HeLa, Jurkat, and COS-1 cell lysates. Flow Cyt: K562. IHC-P: Human kidney tissue.

  • 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

Applications

Our Abpromise guarantee covers the use of ab133493 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 1/10000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 131 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/500 - 1/1000.
IP 1/10 - 1/100.
Flow Cyt 1/200.

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 (By similarity). Plays a central role in microtubule-dependent cell motility via deacetylation of tubulin.

  • Sequence similarities

    Belongs to the histone deacetylase family. HD type 2 subfamily.
    Contains 1 UBP-type zinc finger.

  • Post-translational
    modifications

    Ubiquitinated. Its polyubiquitination however does not lead to its degradation.
    Sumoylated in vitro.

  • Cellular localization

    Nucleus. Cytoplasm. It is mainly cytoplasmic, where it is associated with microtubules.
  • Information by UniProt

  • Database links

    • Alternative names

      • CPBHM antibody
      • FLJ16239 antibody
      • HD 6 antibody
      • HD6 antibody
      • HDAC 6 antibody
      • HDAC6 antibody
      • HDAC6_HUMAN antibody
      • Histone deacetylase 6 (HD6) antibody
      • Histone deacetylase 6 antibody
      • JM 21 antibody
      • JM21 antibody
      • KIAA0901 antibody
      • OTTHUMP00000032398 antibody
      • OTTHUMP00000197663 antibody
      • PPP1R90 antibody
      • Protein phosphatase 1 regulatory subunit 90 antibody
      see all

    Images

    • Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      All lanes : Anti-HDAC6 antibody [EPR1698(2)] (ab133493) at 1/10000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : HDAC6 knockout HeLa cell lysate
      Lane 3 : Wild-type HAP1 cell lysate
      Lane 4 : HDAC6 knockout HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 131 kDa
      Observed band size: 160 kDa
      why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab133493 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab133493 Anti-HDAC6 antibody [EPR1698(2)] was shown to specifically react with HDAC6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264804 (knockout cell lysate ab257145) was used. Wild-type and HDAC6 knockout samples were subjected to SDS-PAGE. ab133493 and Anti-GAPDH antibody [6C5] - Loading Control 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-HDAC6 antibody [EPR1698(2)] (ab133493)
      Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      Lanes 1-2 : Anti-HDAC6 antibody [EPR1698(2)] (ab133493) at 1/10000 dilution
      Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution


      Lanes 1 & 3 & 5 : Wild-type HAP1 cell lysate at 20 µg
      Lanes 2 & 4 : HDAC6 knockout HAP1 cell lysate at 20 µg
      Lane 6 : HDAC6 knockout HAP1 cell lysate

      Predicted band size: 131 kDa



      Lanes 1 and 2: Green signal from target – ab133493 observed at 160 kDa
      Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
      Lanes 5 and 6: Merged (red and green) signal

      ab133493 was shown to specifically react with HDAC6 when HDAC6 knockout samples were used. Wild-type and HDAC6 knockout samples were subjected to SDS-PAGE. ab133493 and ab8226 (loading control to beta actin) were diluted 1/10 000 and 1/1000 respectively and incubated overnight at 4ºC. 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 1/10 000 dilution for 1 h at room temperature before imaging.

    • Flow Cytometry - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      Flow Cytometry - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

      ab133493 staining HDAC6 in K562 (human chronic myelogenous leukemia) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black)

      Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    • Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      All lanes : Anti-HDAC6 antibody [EPR1698(2)] (ab133493) at 1/10000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : K562 cell lysate
      Lane 4 : COS-1 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 131 kDa
      Observed band size: 160 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

      Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling HDAC6 with ab133493 antibody at a dilution of 1/50.

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

    References

    Publishing research using ab133493? Please let us know so that we can cite the reference in this datasheet.

    ab133493 has been referenced in 3 publications.

    • Wattin M  et al. Modulation of Protein Quality Control and Proteasome to Autophagy Switch in Immortalized Myoblasts from Duchenne Muscular Dystrophy Patients. Int J Mol Sci 19:N/A (2018). PubMed: 29316663
    • Phadwal K  et al. Spermine increases acetylation of tubulins and facilitates autophagic degradation of prion aggregates. Sci Rep 8:10004 (2018). PubMed: 29968775
    • Li A  et al. Histone deacetylase 6 regulates the immunosuppressive properties of cancer-associated fibroblasts in breast cancer through the STAT3-COX2-dependent pathway. Oncogene N/A:N/A (2018). PubMed: 29980788

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    Western blot abreview for Anti-HDAC6 antibody [EPR1698(2)]

     Average
    Abreviews
    Application
    Western blot
    Sample
    Pig Cell lysate - whole cell (Heart)
    Gel Running Conditions
    Reduced Denaturing (7,5)
    Loading amount
    20 µg
    Specification
    Heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

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    Verified customer

    Submitted Oct 23 2014

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