Overview

  • Product name

  • Description

    Goat polyclonal to EHD2
  • Host species

    Goat
  • Specificity

    This antibody is expected to recognise EHD1 protein as well as EHD2.
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, ELISA, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Dog
  • Immunogen

    Synthetic peptide:

    C-RLVPPSKRRHKGSA

    , corresponding to C terminal amino acids 529 - 542 of Human EHD2.

  • Positive control

    • Human heart lysate. Lysate prepared from transfected HEK293 transiently expressing EHD2. IF/ICC: A549 cell line.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 0.5% BSA, Tris buffered saline. pH 7.3
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab23935 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 Use a concentration of 10 µg/ml. PubMed: 20008164
Flow Cyt Use a concentration of 10 µg/ml.
ELISA 1/64000. Peptide ELISA
WB Use a concentration of 0.003 - 0.01 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 61 kDa).

Target

  • Function

    Plays a role in membrane reorganization in response to nucleotide hydrolysis. Binds to liposomes and deforms them into tubules. Plays a role in membrane trafficking between the plasma membrane and endosomes. Important for the internalization of GLUT4. Required for normal fusion of myoblasts to skeletal muscle myotubes. Binds ATP; does not bind GTP.
  • Tissue specificity

    Highly expressed in heart and moderately expressed in placenta, lung, and skeletal muscle.
  • Sequence similarities

    Contains 1 EF-hand domain.
    Contains 1 EH domain.
  • Domain

    The EH domain interacts with Asn-Pro-Phe (NPF) motifs of target proteins.
  • Cellular localization

    Cell membrane. Endosome membrane. Colocalizes with GLUT4 in intracellular tubulovesicular structures that are associated with cortical F-actin.
  • Information by UniProt
  • Database links

  • Alternative names

    • EH domain containing 2 antibody
    • EH domain containing 3 antibody
    • EH domain containing protein 2 antibody
    • EH domain-containing protein 2 antibody
    • EHD 2 antibody
    • Ehd2 antibody
    • EHD2_HUMAN antibody
    • EHD3 antibody
    • FLJ96617 antibody
    • PAST 2 antibody
    • PAST homolog 2 antibody
    • PAST2 antibody
    see all

Images

  • Immunofluorescence analysis of paraformaldehyde fixed A431 cells, permeabilized with 0.15% Triton. Primary incubation with ab23935 (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml), showing plasma membrane staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml).

  • Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml), showing nuclear and cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (2 μg/ml).

  • Flow cytometric analysis of paraformaldehyde fixed A431 cells (blue line), permeabilized with 0.5% Triton. Primary incubation ab23935 (10 μg/ml) followed by Alexa Fluor 488 secondary antibody (1 μg/ml). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

  • Anti-EHD2 antibody (ab23935) at 0.3 µg/ml + Human lung lysate at 35 µg

    Predicted band size: 61 kDa

  • Lane 1 : Anti-EHD2 antibody (ab23935) at 0.1 µg/ml
    Lane 2 : Anti-EHD2 antibody (ab23935) at 0.3 µg/ml

    Lane 1 : A549 cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 61 kDa

  • ICC/IF image of ab23935 stained A549 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA  / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23935, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96931, DyLight® 488 donkey anti-goat IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM

  • Anti-EHD2 antibody (ab23935) at 0.003 µg/ml + Human Heart lysate (RIPA buffer, 35µg total protein per lane).

    Predicted band size: 61 kDa
    Observed band size: 61 kDa



    Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
  • Anti-EHD2 antibody (ab23935) at 0.1 µg/ml + Human Placenta lysate in RIPA buffer at 35 µg

    Predicted band size: 61 kDa

References

This product has been referenced in:

  • Keshavarz M  et al. Caveolin-3 differentially orchestrates cholinergic and serotonergic constriction of murine airways. Sci Rep 8:7508 (2018). Read more (PubMed: 29760450) »
  • Mendoza-Topaz C  et al. BioID identifies proteins involved in the cell biology of caveolae. PLoS One 13:e0209856 (2018). Read more (PubMed: 30589899) »
See all 18 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question
Answer

We have not tested this antibody with hamster, only human and mouse. The immunogen used to raise this antibody is RLVPPSKRRHKGSA and is 100% homologous with Chinese Hamster EDH1 (note that this antibody should recognize both EDH1 and EDH2).  Therefore, we would expect this antibody to cross-react with Chinese Hamster, but we couldn't guarantee it as this hasn't been tested.

Read More
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa)
Total protein in input
300 µg
Specification
HeLa
Immuno-precipitation step
Protein A

Debora Rapaport

Verified customer

Submitted Feb 18 2011

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