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Polyacrylamide gels shrink during staining, so comparison of an immunologically probed membrane to a stained gel is not practical. To determine the exact location of a specific antigen in relation to other proteins, compare two blotted
Tris–tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)wasperformedusingMiniProtean II cell (BioRad, Sunnyvale, CA, USA) according to protocol of Scha¨gger and von Jagov [54]. After separation, the gels were fixed by
