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| Hi, You're right - better to describe the already tested methods. The methods we have tried are: 1. 1% SDS in PBS overnight at 37degreesC. This has variable results depending on the antibody, but doesn't result in much loss of protein. 2. 2% SDS, 100mM beta-mercaptoethanol, 62.5mM Tris pH 7.4, for 30 minutes either at 50 degreesC or at room temperature. This works for blots of cultured cell lysates, but results in high loss of protein so it is too harsh for blots of embryo protein. We're looking at the degree of phophorylation of several proteins, so we need to have good removal of antibody, but we have very little protein, so we need to minimize protein loss. Thanks, Ann |
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| blots , stripping , western |
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