Validated using a knockout cell line
Recombinant
RabMAb

Anti-Rab4 antibody [EPR3042] - Early Endosome Marker (ab108974)

Overview

  • Product name
    Anti-Rab4 antibody [EPR3042] - Early Endosome Marker
    See all Rab4 primary antibodies
  • Description
    Rabbit monoclonal [EPR3042] to Rab4 - Early Endosome Marker
  • Tested applications
    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human Rab4 aa 1-100.

  • Positive control
    • MCF7, PC12, Neuro-2a, mouse brain, rat brain and fetal brain cell lysates
  • 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 ab108974 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 1/2000 - 1/10000. Detects a band of approximately 25 kDa (predicted molecular weight: 24 kDa).
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for Flow Cyt or IHC-P.
  • Target

    Images



    • Predicted band size : 24 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Rab4 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Human fetal brain cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab108974 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Unpurified ab108974 was shown to recognize Rab4 when Rab4 knockout samples were used, along with additional cross-reactive bands. Wild-type and Rab4 knockout samples were subjected to SDS-PAGE. Unpurified ab108974 and ab8245 (loading control to GAPDH) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • All lanes : Anti-Rab4 antibody [EPR3042] - Early Endosome Marker (ab108974) at 1/2000 dilution (purified)

      Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
      Lane 3 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
      Lane 4 : Mouse brain lysates
      Lane 5 : Rat brain lysates

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size : 24 kDa
      Observed band size : 25 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Rab4 with purified ab108974 at 1:100 dilution (8.8μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunofluorescent staining of Rab4 in HeLa cells, using unpurified ab108974 at a 1/100 dilution.

    • All lanes : Anti-Rab4 antibody [EPR3042] - Early Endosome Marker (ab108974) at 1/2000 dilution (unpurified)

      Lane 1 : MCF7 cell lysates
      Lane 2 : PC12 cell lysates
      Lane 3 : Fetal brain cell lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 24 kDa
      Observed band size : 25 kDa (why is the actual band size different from the predicted?)

    References

    ab108974 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Deux anticorps semblent intéressants pour la détection de Rab4 chez la Drosophile:
    Le premier est un polyclonal de lapin dont l'immunogène présente 86% d'homologie avec la protéine d'intéret (voir fichier pdf ci-joint "ab78970 vs Drosophila"). La r...

    Read More

    Thank you for getting back to me with that information.

    I am sorry, I did miss understand you, I thought you were using tissue, not cell lines. This does make the band at 50 kDa unexpected, although I wouldsay that a"no-primary" con...

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    Thank you for the swift reply.

    The extra details have made it much easier to understand the problems you have been encountering. Although there is a faint band at the expected size (˜25 kDa), as you have described, the non-specific band at ˜4...

    Read More

    Thank you for contacting us.

    I am sorry to hear you are experiencing difficulties with the anti-Rab4 antibody (ab108974). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. ...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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