I am doing imaging of OCT-1, and although my secondary antibody is correct, I am getting non specific binding, showing in negative control (no primary AB). Would this be related more to my 2AB or maybe attempt a different blocking buffer?
Sang Ho Lee
Hello Samulel.
If you did everything properly, you still get the same results. Then, your 1st and 2nd abs have gone off, if you directly prepared with carrier protein (BSA), aliquoted, stored at -80C, thawed, centrifuged and used it. Get new antibodies from a company not to waste many months.
Regards.
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Nora Fekete-Drimusz
Assuming your antibodies haven't gone off, have you tried decreasing the concentration of the secondary antibody?
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