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Please find attached images/comments about our optimization for western blots using your products. We have not been able to produce satisfactory results with the following antibodies: ab2500, ab6586, ab7033, ab1157 We also have questions pertaining to the following antibodies: ab1828, ab3644. We are happy to receive advice from past experiences that Abcam may have had in the process of testing these antibodies and are open to continuing the optimization process. However, if satisfactory results are not obtained, we will count on the Abcam product guarantee.
Asked on Jun 22 2006
Thank you for your enquiry. I am sorry to hear that you have been having difficulties with these antibodies. The results that you have been obtaining are certainly troubling. What concerns me the most are the two bands that you are consistently detecting at approximately 25 and 37KDa. You are consistently detecing cross reacting bands and never detecting the endogenous target. Whilst some of the antibodies that you detail have not been tested using rat samples; potentially explaining the absence of reactivity with the endogenous band the presence of the two bands at the aforementioned molecular weight accross many of your samples seems to suggest problems with the secondary antibody. Especially in view of the fact that you have changed this antibody and are in fact using three independent secondary antibodies: Goat polyclonal to Rabbit IgG H&L (HRP) (ab6721) Goat polyclonal to Rabbit IgG + Mouse IgG & IgM - prediluted (HRP polymer) (ab2891) Rabbit polyclonal to Goat IgG H&L (HRP) (ab6741) I would appreciate it if you could provide me with protocol details of the approach that you are using so that I can better determine potential reasons for these reacting bands. We have not observed such reactivity with any of the antibodies you list. I am particularly interested in the generic protocol that you have been applying. To provide me with these details please click on the link below and complete our on line technical questionaire. If you have modified the protocol for any of the antibodies at any point please mention this. I am also very interested in the method of sample preparation and the control antibody (loading control) that you have used. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=2500&mode=questionaire I look forward to hearing from you.
Answered on Jun 28 2006