Overview

  • Product name

    Anti-ENO1 antibody [EPR19758]
    See all ENO1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19758] to ENO1
  • Host species

    Rabbit
  • Specificity

    ab227978 does not cross-react with ENO2 or ENO3.
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ENO1 aa 50-150. The exact sequence is proprietary.
    Database link: P06733

  • Positive control

    • WB: MCF7, RAW 264.7, Jurkat, HeLa, NIH/3T3 and C6 whole cell lysates; human skeletal muscle tissue lysate; mouse brain, heart and kidney tissue lysates; rat brain, heart, liver and spleen tissue lysates. IP: Jurkat whole cell lysate. IHC-P: Human pancreas and clear cell renal cell carcinoma tissue; mouse and rat pancreas tissue. ICC/IF: HeLa and NIH/3T3 cells.
  • 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.

Applications

Our Abpromise guarantee covers the use of ab227978 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
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 47 kDa.
ICC/IF 1/500.
Flow Cyt 1/600.
IP 1/30.

Target

  • Function

    Multifunctional enzyme that, as well as its role in glycolysis, plays a part in various processes such as growth control, hypoxia tolerance and allergic responses. May also function in the intravascular and pericellular fibrinolytic system due to its ability to serve as a receptor and activator of plasminogen on the cell surface of several cell-types such as leukocytes and neurons. Stimulates immunoglobulin production.
    MBP1 binds to the myc promoter and acts as a transcriptional repressor. May be a tumor suppressor.
  • Tissue specificity

    The alpha/alpha homodimer is expressed in embryo and in most adult tissues. The alpha/beta heterodimer and the beta/beta homodimer are found in striated muscle, and the alpha/gamma heterodimer and the gamma/gamma homodimer in neurons.
  • Pathway

    Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 4/5.
  • Sequence similarities

    Belongs to the enolase family.
  • Developmental stage

    During ontogenesis, there is a transition from the alpha/alpha homodimer to the alpha/beta heterodimer in striated muscle cells, and to the alpha/gamma heterodimer in nerve cells.
  • Post-translational
    modifications

    ISGylated.
  • Cellular localization

    Nucleus and Cytoplasm. Cell membrane. Cytoplasm > myofibril > sarcomere > M line. Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form. ENO1 is localized to the M line.
  • Information by UniProt
  • Database links

  • Alternative names

    • 2 phospho D glycerate hydro lyase antibody
    • 2-phospho-D-glycerate hydro-lyase antibody
    • Alpha enolase antibody
    • Alpha enolase like 1 antibody
    • Alpha-enolase antibody
    • C myc promoter binding protein antibody
    • C-myc promoter-binding protein antibody
    • EC 4.2.1.11 antibody
    • eno1 antibody
    • ENO1L1 antibody
    • ENOA_HUMAN antibody
    • Enolase 1 (alpha) antibody
    • Enolase 1 (alpha) like 1 antibody
    • Enolase 1 antibody
    • Enolase alpha antibody
    • MBP 1 antibody
    • MBP-1 antibody
    • MBP1 antibody
    • MBPB1 antibody
    • MPB 1 antibody
    • MPB-1 antibody
    • MPB1 antibody
    • MYC promoter binding protein 1 antibody
    • NNE antibody
    • Non neural enolase antibody
    • Non-neural enolase antibody
    • Phosphopyruvate hydratase antibody
    • Plasminogen binding protein antibody
    • Plasminogen-binding protein antibody
    • PPH antibody
    • Tau crystallin antibody
    see all

Images

  • All lanes : Anti-ENO1 antibody [EPR19758] (ab227978) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Mouse heart tissue lysate
    Lane 3 : Mouse kidney tissue lysate
    Lane 4 : Rat brain tissue lysate
    Lane 5 : Rat heart tissue lysate
    Lane 6 : Rat liver tissue lysate
    Lane 7 : Rat spleen tissue lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldegyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 with ab227978 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in HeLa cell line. 

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

    -ve control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution .

  • Immunohistochemical analysis of paraffin-embedded human clear cell renal cell carcinoma tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and nuclear staining in human clear cell kidney carcinoma (PMID: 26037892) is observed. Counter stained with hematoxylin.

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

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 with ab227978 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).

    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

  • ENO1 was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab227978 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227978 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

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

    Lane 2: ab227978 IP in Jurkat whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227978 in Jurkat whole cell lysate.

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

    Exposure time: 3 seconds.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of human pancreas (PMID: 19425054) is observed. Counter stained with hematoxylin.

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

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of mouse pancreas is observed. Counter stained with hematoxylin.

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

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of rat pancreas is observed. Counter stained with hematoxylin.

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

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-ENO1 antibody [EPR19758] (ab227978) at 1/1000 dilution

    Lane 1 : Human skeletal muscle tissue lysate at 20 µg
    Lane 2 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
    Lane 5 : C6 (rat glial tumor glial cell line) whole cell lysate at 10 µg
    Lane 6 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
    Lane 7 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg

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

    Developed using the ECL technique.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lane 1: 15 seconds.

    Lanes 2-7: 1 second.

  • Immunofluorescent analysis of 4% paraformaldegyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling ENO1 with ab227978 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in NIH/3T3 cell line.

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

    -ve control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluo 488) (ab150077) secondary antibody at 1/1000 dilution .

References

ab227978 has not yet been referenced specifically in any publications.

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