Overview

  • Product name

    Anti-HDAC1 + HDAC2 antibody [EPR20327]
    See all HDAC1+HDAC2 primary antibodies
  • Description

    Rabbit monoclonal [EPR20327] to HDAC1 + HDAC2
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human HDAC2 aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: Q92769

  • Positive control

    • WB: His-tagged human HDAC2 (aa339-488) and HDAC1 (aa1-482) recombinant proteins; HeLa, LNCaP, SH-SY5Y, 293, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; human fetal heart and fetal kidney lysates; mouse and rat brain and spleen lysates; rat kidney lysates. IHC-P: Human testis, tonsil, prostate hyperplasia, prostate cancer, breast cancer and synovial sarcoma tissues; Mouse colon tissue. ICC/IF: HEK-293 and NIH/3T3 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab219054 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
ICC/IF 1/1000.
IP 1/30.
WB 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/700.

Target

Images

  • All lanes : Anti-HDAC1 + HDAC2 antibody [EPR20327] (ab219054) at 1/10000 dilution

    Lane 1 : His-tagged human HDAC2 recombinant protein (aa339-488)
    Lane 2 : His-tagged human HDAC1 recombinant protein (aa1-482)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 55 kDa
    Observed band size: 22,60 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on human testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HEK-293 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-HDAC1 + HDAC2 antibody [EPR20327] (ab219054) at 1/2000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : LNCaP (Human prostate cancer cell line) whole cell lysate
    Lane 3 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
    Lane 4 : 293 (Human epithelial cell line from embryonic kidney) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-HDAC1 + HDAC2 antibody [EPR20327] (ab219054) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-HDAC1 + HDAC2 antibody [EPR20327] (ab219054) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse spleen lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat kidney lysate
    Lane 5 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/3-5: 3 minutes; Lane 2: 10 seconds.

  • All lanes : Anti-HDAC1 + HDAC2 antibody [EPR20327] (ab219054) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on lymphocytes of human tonsil is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human prostate hyperplasia tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on luminal epithelial cells of human prostate hyperplasia, but negative staining on basal cells.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on tumor cells of prostate cancer; weak or negative staining on basal cells.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on tumor cells of human breast cancer is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human synovial sarcoma tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on human synovial sarcoma is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Nuclear staining on mouse colon is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HDAC1 + HDAC2 with ab219054 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HDAC1 + HDAC2 with ab219054 at 1/700 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • HDAC1 + HDAC2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab219054 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab219054 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10 µg (Input).

    Lane 2: ab219054 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219054 in HeLa whole cell lysate.

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

    Exposure time: 1 second.

References

This product has been referenced in:

  • Liao M  et al. LncRNA EPB41L4A-AS1 regulates glycolysis and glutaminolysis by mediating nucleolar translocation of HDAC2. EBioMedicine 41:200-213 (2019). Read more (PubMed: 30796006) »
See 1 Publication for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab219054.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up