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Novel Fluorescence Labeling and High-Throughput Assay Technologies for in Vitro Analysis of Protein Interactions

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Protein samples were separated by 412% SDS-PAGE, transferred onto PVDF membranes (GE Healthcare, Milan, Italy), and incubated with antibodies targeting and Alpha-Smooth Muscle Actin (catalog n A2547, Sigma-Aldrich)

Extending the treatment time to 3 min as well as to 5 min caused no changes in the fragmentation pathway

But if you happen to come across moldy, shriveled, or discolored shells (theyre usually a deep brown), toss them