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What dilutions of ab8976 have you tried? 1:500, 1:400, 1:300 Do you know that the secondary antibody is working properly? Yes, we used them to detect alpha actin, GAPDH and some other antibodies in the same days/same membranes. Also, did you ensure that the protein transferred properly to the membrane? Yes, we detect GAPDH on the same membrane. Thank you again, and i look forward to hearing from you.
Asked on Aug 10 2005
Thank you for your email. I would expect for you to detect desmin in your samples, smooth muscle cells, and you certainly have tried high enough concentrations of ab8976. How long did you incubate with the primary? I would suggest incubating overnight at 4C. Other than that, your protocol looks fine to me. We have not had any other complaints regarding this product, and there is a possibilty that the vial you received had gone off during the shipping process. I can offer to send you a free of charge replacement vial of ab8976 or can offer you a credit note or refund. Please let me know which you would prefer.
Answered on Aug 11 2005