02 February 2020 3 6K Report

Hello,

I am wondering if it is alright to reuse my antibodies for immunofluorescence experiments. I have reused primary antibodies with success for the first two experiments, but then I saw a diminished signal upon the third use.

Is it alright to reuse antibodies in this way? Is it true that antibodies may form complexes with other molecules during incubation which can then stick during the next experiment and possibly cause a false positive? How about secondary antibodies? Can they form complexes with residual primary antibodies in suspension which can then pass on to the next experiment? What is the biggest risk of reusing antibodies for immunofluorescence?

For immunofluorescence, I dilute my antibodies in blocking buffer composed of 10% FBS and 2% BSA in PBS with 10mM sodium azide (NaN3).

Thank you!

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