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

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 référence de cet anticorps est: ab78970 et le lien vers la fiche technique est www.abcam.com/ab78970.
Le second est un RabMAb, ou monoclonal de lapin, dont l'immunogène présente 71% d'homologie avec la protéine d'intéret (voir fichier pdf ci-joint "ab108974 vs Drosophila"). La référence de cet anticorps est: ab108974 et le lien vers la fiche technique est www.abcam.com/ab108974.
Nous avons en ce moment une promotion qui vous donne droit à un monoclonal de lapin gratuit pour l'achat d'un autre anticorps.
De plus, je peux vous offrir une offre de test pour le ab78970. En voici les détails:
1. Nous confirmer que vous souhaitez tester ab78970 chez la drosophile afin de recevoir le code de remise correspondant. Ce code doit être édité avant l’achat de ab78970
2. Acheter ab78970 par téléphone, fax ou internet (www.abcam.com)
3. Le tester chez la drosophile
4. Nous faire part de vos résultats, positifs ou négatifs, grâce à notre système Abreview. Pour plus d’informations https://www.abcam.com/abreviews
5. Après soumission de votre Abreview, appeler notre service clientèle afin de passer votre prochaine commande grâce au code de réduction. Ce code de réduction est utilisable 120 jours après son édition et utilisable lors de l’achat d’un autre anticorps primaire.
Même si ab78970 ne pas fonctionne pas chez la drosophile, après soumission de votre Abreview, vous recevrez quand même un avoir de la valeur de l'anticorps testé.
Termes et Conditions : www.abcam.com/collaborationdiscount.
Merci de m'indiquer si vous souhaitez bénéficier de cette offre de test pour le ab78970. En attendant votre réponse je vous envoie dans un autre email un devis pour le ab78970 et le ab108974.

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" control would still be worth performing.

I am sorry to hear that two of our other antibodies have not been performing as expected either. Could you please provide information as to what results you have had with these antibodies? I would really like to look into this further in order to understand what may be going wrong. If possible, could you fill out the questionnaire I have attached to this email, adding any images that you have of the blots would also be very useful.

Could you also please provide mewith the catalogue numbers and lot numbers of the antibodies used? If youdo not have access to this then the order numbers?Which is the new monoclonal you have purchased from us?

May I alsoask, have you used any other anti-Rab4 antibodies from any other suppliers? What have been the results of this?

I look forward to receiving your reply.

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 ˜45-50 kDa is much stronger. I think this band may possible be from the rat IgG being picked up but the goat anti-rabbit secondary antibody. Could you giveme further detailsof the secondary antibody used? Has this secondary been used with rat samples previously? I would suggest thatit may be worthwhile toperforma no-primary control and/or use adifferent anti-rabbit antibodyin order to investigate this.

The blot produced, is itthe first time ithas been probed or has a different primary antibody beendetected using the same blot followed by stripping? If so couldyou describe how this was performed?

Many thanks for your continued cooperation.

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.

Thank you forproviding the details of the experiment you have performed. This information will allow us to investigate this case internally and initiate additional testing where necessary. In order to understand the situation more fully, and hopefully help to improve the results obtained so far, could you please provide the following additional information:

1. What lysateshave been used with the antibody and how have these been prepared prior to loading on the gel? - how was the sample lysed (lysis buffer, protease inhibitors?) and was the sample denatured by boiling and a reducing agent used?

2. Was any loading control preformed in order to ascertain the quality of the samples? I.e. detection of something like actin or tubulin?

3. Has the secondary antibody been used successfully before in Western blotting with any other primary antibodies?

4. If you would be able to send an image of the blot obtained this would be very helpful in understanding what may be contributing to the unexpected results.

5. Would you be able to tell be the order number from which you ordered this antibody?

If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.