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WebGenerally, 4°C overnight will give more efficient and specific staining. However, many antibodies will also work very well with one or two hours incubation at room temperature. WebIt depends on the binding affinity of the antibody to the target protein, usually 2 hours at room temperature incubation or 4°C incubation overnight. After incubating with the primary antibody, the membrane is then washed with TBST or PBST buffer for 4-5 times, about 5 minutes each time with agitation, to remove residual unbound primary antibody. health informatics salary bls

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WebHi, blocking overnight at 4C basically has the same effect as 1-2 hours at RT, so it should make no difference. The incubation time of the first AB is dependent on how good the AB … http://mrcanu.pharm.ox.ac.uk/sites/default/files/pdfs/somogyitakagi1982neuroscience.pdf WebGenerally, 4°C overnight will give more efficient and specific staining. However, many antibodies will also work very well with one or two hours incubation at room temperature. health informatics scholarships for women