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A Customer has sent me an e-mail claiming about a product bought by him. According to the information given, he acquired an antibody, ab97264 and it has been applied in flow cytometry. However, he has obtained unspecific signal, that means, he obtained positive signal in absence of the antibody, using the secondary antibody in different dilutions: 1/50 and 1/25. First, the customer though that could be a blocking problem, so he assayed different dilutions with BSA (1-10%), but he got the same negative results. The assays had been performed in non-permeabilized cells, so in absence of detergents and without fixation solution at 4ºC. I will appreciate you may assistance in this question as soon as possible. Best regards
Asked on Sep 14 2011
Thank you for your enquiry. I would recommend that the customer increase the dilition factor for this secondary antibody from 1:50 to 1:200. I believe this will reduce the background. I hope this is helpful. Please let me know if there are any further questions.
Answered on Sep 14 2011