Recombinant
RabMAb

Recombinant Anti-EHD1 antibody [EPR4954] (ab109311)

Submit a review Q&A (1)References (13)

Overview

  • Product name

    Anti-EHD1 antibody [EPR4954]
    See all EHD1 primary antibodies

  • Description

    Rabbit monoclonal [EPR4954] to EHD1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human EHD1. The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, Human testis, SW480, SK-OV-3, Human brain, HeLa, Mouse brain, and Rat brain lysates. IHC-P: human thyroid carcinoma, kidney and tonsil, mouse lung, and rat lung tissues. IP: HeLa 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109311 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 1/1000 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 61 kDa).
IP 1/10 - 1/100.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

(heat to 98°C, allow to cool for 10-20 minutes)

1:120 in human tissues / 1:1600 in mouse and rat tissues

See IHC antigen retrieval protocols.’

Target

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling EHD1 with purified ab109311 at 1/120 dilution (2.85 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-EHD1 antibody [EPR4954] (ab109311)
    Western blot - Anti-EHD1 antibody [EPR4954] (ab109311)
    All lanes : Anti-EHD1 antibody [EPR4954] (ab109311) at 1/10000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 61 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?

  • Immunoprecipitation - Anti-EHD1 antibody [EPR4954] (ab109311)
    Immunoprecipitation - Anti-EHD1 antibody [EPR4954] (ab109311)

    ab109311 (purified ) at 1/30 dilution (2ug) immunoprecipitating EHD1 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab109311 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109311 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat lung tissue sections labeling EHD1 with purified ab109311 at 1/1600 dilution (0.21 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-EHD1 antibody [EPR4954] (ab109311)
    Western blot - Anti-EHD1 antibody [EPR4954] (ab109311)
    Anti-EHD1 antibody [EPR4954] (ab109311) at 1/10000 dilution (Purified) + Human brain lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 61 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-EHD1 antibody [EPR4954] (ab109311)
    Western blot - Anti-EHD1 antibody [EPR4954] (ab109311)
    All lanes : Anti-EHD1 antibody [EPR4954] (ab109311) at 1/1000 dilution ((unpurified))

    Lane 1 : HepG2 cell lysates
    Lane 2 : Human testis lysates
    Lane 3 : SW480 cell lysates
    Lane 4 : SK-OV-3 cell lysates
    Lane 5 : HeLa cell lysates

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 61 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse lung tissue sections labeling EHD1 with purified ab109311 at 1/1600 dilution (0.21 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)

    Immunohistochemical analysis of EHD1 in paraffin embedded Human kidney tissue, using unpurified ab109311 at a 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHD1 antibody [EPR4954] (ab109311)

    Immunohistochemical analysis of EHD1 in paraffin embedded Human tonsil tissue, using unpurified ab109311 at a 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Kamerkar SC  et al. A Screen for Membrane Fission Catalysts Identifies the ATPase EHD1. Biochemistry 58:65-71 (2019). Read more (PubMed: 30403133) »
  • Insinna C  et al. Investigation of F-BAR domain PACSIN proteins uncovers membrane tubulation function in cilia assembly and transport. Nat Commun 10:428 (2019). Read more (PubMed: 30683896) »
See all 13 Publications for this product

Publishing research using ab109311? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Question

I would like to use this Ab to detect EHD1 on a western blot. Could you tell me the peptide it was rasised against as I would like to ensure that it is in a region not present in EHD2, 3, and 4. Thank you.

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Answer

Thank you for contacting Abcam regarding our EHD1 antibody, ab109311. I have confirmed with the laboratory that this antibody is not predicted to cross react with other EHD proteins EHD2, 3, and 4. Sequence analysis shows that the immunogen shares no significant sequence identity with them. It shares 69% identity with EHD3. I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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