m*t 发帖数: 1 | 1 Let me make a bold guess just for fun. :)))
Assume that the band you have seen after 2Ab is the correct band instead of a
crossreactivity artifact, I guess what was happening is that you 1Ab has a
very weak interaction against the fusion protein. The Ab can be easily washed
off in the subsequent washing steps, however, when you added destain solution,
the methanol fixes the 1Ab on the blot so that it woon't be totally washed.
Therefore the 2Ab can detect the 1Ab on the membrane, and that's also |
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