Recombinant
RabMAb

Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (ab189891)

Overview

  • Product name
    Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407]
    See all ENO1 + ENO2 + ENO3 primary antibodies
  • Description
    Rabbit monoclonal [EPR18407] to ENO1 + ENO2 + ENO3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ENO1 + ENO2 + ENO3 aa 250-350. The exact sequence is proprietary.
    Database link: P13929

  • Positive control
    • WB: Human ENO1, ENO2 and ENO3 full length recombinant proteins; Human fetal heart, fetal kidney and fetal spleen lysates; HeLa, Jurkat, MCF7, A431, C6, RAW 264.7 and NIH/3T3 whole cell lysates; Mouse brain and heart lysates; Rat brain and heart lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab189891 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
Flow Cyt 1/70.
ICC/IF 1/500.
WB 1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).

Target

  • Relevance
    Enolase 1 is a 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. Enolase 2 has neurotrophic and neuroprotective properties on a broad spectrum of central nervous system (CNS) neurons. Binds, in a calcium-dependent manner, to cultured neocortical neurons and promotes cell survival. Enolase 3 appears to have a function in striated muscle development and regeneration.
  • Cellular localization
    ENO1: Cytoplasm. Cell membrane. Cytoplasm, myofibril, sarcomere, M-band. Note: Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form. ENO1 is localized to the M-band. ENO2: Cytoplasm. Cell membrane. Note: Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form ENO3: Cytoplasm. Note: Localized to the Z line. Some colocalization with CKM at M-band.
  • Database links
  • Alternative names
    • 2-phospho-D-glycerate hydro-lyase antibody
    • Alpha-enolase antibody
    • Beta-enolase antibody
    • C-myc promoter-binding protein antibody
    • Crystallin Tau, included antibody
    • ENO1L1 antibody
    • Enolase 1, (alpha) antibody
    • Enolase 2 (gamma, neuronal) antibody
    • Enolase 2 antibody
    • Enolase 3 (beta, muscle) antibody
    • Enolase 3 antibody
    • Enolase alpha antibody
    • Enolase, beta antibody
    • Enolase, gamma antibody
    • Enolase, muscle specific antibody
    • Enolase, neuron specific antibody
    • Enolase, nonneuronal, included antibody
    • Gamma-enolase antibody
    • GSD13 antibody
    • MBP-1 antibody
    • MPB1 antibody
    • MSE antibody
    • Muscle-specific enolase antibody
    • Neural enolase antibody
    • Neuron-specific enolase antibody
    • NNE antibody
    • Non-neural enolase antibody
    • NSE antibody
    • Phosphpyruvate hydratase antibody
    • Plasminogen-binding protein antibody
    • Skeletal muscle enolase antibody
    see all

Images

  • All lanes : Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (ab189891) at 1/1000 dilution

    Lane 1 : Human ENO1 full length recombinant protein
    Lane 2 : Human ENO2 full length recombinant protein
    Lane 3 : Human ENO3 full length recombinant protein

    Lysates/proteins at 0.02 µg per lane.

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

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


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human ENO1 full length recombinant protein (Cat#:ab89248) contains aa1-434. Human ENO2 full length recombinant protein contains aa1-434 with a His-Tag®.  Human ENO3 full length recombinant protein(Cat#:ab113127) contains aa1-434 with a His-Tag®. Human ENO2 full length recombinant protein was made in-house.

  • All lanes : Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (ab189891) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time : Lane 1 and 2: 5 seconds; Lane 3: 15 seconds.

  • All lanes : Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (ab189891) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 4 : A431 (Human epidermoid carcinoma cell line) 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: 47 kDa
    Observed band size: 47 kDa


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Lanes 1-4 : Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (ab189891) at 1/1000 dilution
    Lanes 5-7 : Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (ab189891) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat heart lysate
    Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 6 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 7 : 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 1/10000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time : Lane 1-4: 5 seconds; Lane 5-7: 1 second.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 + ENO2 + ENO3 with ab189891 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing mostly cytoplasmic staining on Hela cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab189891 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ENO1 + ENO2 + ENO3 with ab189891 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing mostly cytoplasmic staining on NIH/3T3 cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody -Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab189891 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line)  labeling ENO1 + ENO2 + ENO3 with ab189891 at 1/70 dilution (red) compared with a Rabbit IgG,monoclonal - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

References

This product has been referenced in:

See 1 Publication for this product

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