Overview

  • Product name

    Anti-Rab5 antibody [3A4]
    See all Rab5 primary antibodies
  • Description

    Mouse monoclonal [3A4] to Rab5
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to amino acids 1-215 of Human Rab5 purified from E. coli.

  • Positive control

    • WB: Extracts of mouse and human brain. ICC/IF: U87MG cells.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 24/01/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab66746 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 Use at an assay dependent concentration. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).
ICC/IF Use at an assay dependent concentration.

Target

Images

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Rab5 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Hu brain whole cell lysate (20 µg)
    Lane 4: Ms brain whole cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab66746 observed at 25 kDa. Red - loading control, ab181602, observed at 37 kDa.

    This image was produced using the ascites version of this antibody. 

  • Immunocytochemistry/ Immunofluorescence analysis of Rab5a in U87MG cells. The cell was stained with ab66746 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
  • Anti-Rab5 antibody [3A4] (ab66746) at 1/500 dilution + mouse brain at 45 µg

    Secondary
    goat anti-mouse secondary antibody conjugated to HRP

    Predicted band size: 24 kDa

  • ab66746 staining Rab5 in the U87MG human cell line at a dilution of 1/100. The cells were fixed before staining with a mixture of acetone and methanol, 1:1. DAPI was used fhor nuclear staining.

    This image was produced using the ascites version of this antibody. 

  • Anti-Rab5 antibody [3A4] (ab66746) at 1/500 dilution + extracts of mouse brain at 45 µg

    Secondary
    HRP-conjugated goat anti-mouse

    Developed using the ECL technique.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa



    This image was produced using the ascites version of this antibody. 

References

This product has been referenced in:

  • Winer H  et al. Autophagy differentially regulates TNF receptor Fn14 by distinct mammalian Atg8 proteins. Nat Commun 9:3744 (2018). Read more (PubMed: 30218067) »
  • Del Cid-Pellitero E  et al. Homeostatic Changes in GABA and Glutamate Receptors on Excitatory Cortical Neurons during Sleep Deprivation and Recovery. Front Syst Neurosci 11:17 (2017). Read more (PubMed: 28408870) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 6% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 09 2016

Answer

Thank you for your email.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab66746.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.


Here is the information about the testing discount code:

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab66746 in ICC/IF. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for ICC/IFand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

I wish you the best of luck with your research.

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Answer

Thank you for contacting us.

Yes, I'd be happy to send you one of the mouse antibodies, but am afraid I cannot make the final decision for you. I'd be able to make a recommendation, though.
Both antibodies (ab66746,ab55657) have not been tested inICC/IF, thus we cannot guarantee either of the antibodies forthisapplication. However, ab66746 was tested in mouse samples in WB. Thus, I would recommend using this antibody.

Since ICC/IF is untested, I would also send you a testing discount code: For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the antibody/protein/kit at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody/protein at the full price (100%) of the antibody/protein you have tested, or a full price discount for the amount paid for the kit. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
This programapplies to ELISA kits and other kits we offer.

Please let me know if I shall go ahead with the free of charge replacement with ab66746 and the testing discount code for trying it in ICC/IF?

I look forward to hear back from you and assist you further.

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