I am screening antibodies and I discovered that one of them is listed on DSHB (Developmental Studies Hybridoma Bank) as being detergent sensitive. I am wondering what would vary from my normal blocking solution, which contains 0.5 % Triton, with a detergent free blocking solution?
Perhaps I am overthinking this but I appreciate any input.
Blocking and permeabilization will be a separate step from antibody addition. I would use your normal permeabilization/blocking solution (with detergent) prior to antibody addition and then use the solution without detergent. I would imagine that as long as you sufficiently wash the sample of detergent and add the antibody in a detergent free solution you will be fine. Does this make sense? If you are mixing your own solutions, this should be easy to do! Let me know if you have more questions.
Elliot
Dear Griffin,
I strongly agree with Elliot's answer. However, I think 0.5% Triton X-100 solution is still higher for the permeabilization step. Before blocking, you may use 0.1-0.25% Triton X-100 and properly washed the sample with PBS (pH 7.4).
Thank you.
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