PROTEIN EXTRACTION AND WESTERN BLOTTING FROM METHACARN-FIXED TISSUE

Polyacrylamide gel electrophoresis (PAGE) of proteins with subsequent western blotting has become a routine technique for the analysis of pr oteins from both cultured cells and fresh whole tissue. We have develo ped a method to extract proteins from methacarn-fixed tissue which ren ders them suitable for SDS-PAGE and western blotting. With a panel of antibodies to specific intermediate filaments, transforming growth fac tor-alpha (TGF-alpha), and albumin, immunohistochemistry was performed in parallel with western blotting on sections cut from methacarn-fixe d samples of normal rat liver and liver from rats treated under a regi me which induces oval cell proliferation. Immunohistochemistry enabled the determination of changes in tissue distribution and abundance of the target proteins, which was mirrored by the corresponding western b lot data. This technique can be especially effective when used in conj unction with immunohistochemistry. Tissue samples are easy to prepare, avoiding the precautions which need to be taken when handling fresh t issue (Abstract: J Pathol 1994; 173S: No. 41).