Application of transfer onto PVDF and nitrocellulose membranes in the purification and structural analyses of proteins
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Replicas of SDS-PAGE gels widen the range of application of electrophoresis to the studies of diverse biological properties of proteins as well as to the elucidation of their structures. Most often proteins are transferred to membranes made of polyvinylidene difluoride (PVDF) or nitrocelulose.Usually, a "semi-dry" blotting apparatus is used for electrophoretic transfer.SDS-PAGE gel replicas are used for the studies of these biological properties which cannot be studied directly on polyacrylamide gels. The major applications of this technique is in the immunoblotting which allows for the antigen-antibody reaction on the membrane.This reaction, after necessary amplification, is visualized with a variety of reagents.Protein microsequencing is another important application of SDS-PAGE gel replicas.In this case, a replica is stained with a general protein stain, and a band of interest is excised and used to obtain partial or complete amino acid sequence in an automatic sequencer.
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