Protein blotting with direct blotting electrophoresis.
Direct blotting electrophoresis, a method designed to be of general application for the separation and electroblotting of macromolecules, has been adapted to produce protein blots suitable for subsequent processing by standard techniques such as dye staining or immunological detection. After their separation in a very short gel the protein bands are electrophoresed out of the gel onto an immobilizing matrix. The matrix which is moved across the bottom of the gel by a conveyor belt binds these proteins with high affinity. Once the protein samples have been loaded onto the gel and electrophoresis has been started, no further intervention is needed until the blot is completed. The total expenditure of time for such a direct blot is less than 4 h for a mixture of proteins in the molecular weight range of 14-70 kDa. The staining sensitivity of directly blotted proteins is about 200 ng protein per band as revealed by India ink staining.
Beck
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Medical Research Council, Laboratory of Molecular Biology, Cambridge, England.
D003101:Collodion; D004396:Coloring Agents; D004586:Electrophoresis / Q000379:methods; D007120:Immunochemistry; D008567:Membranes, Artificial; D008970:Molecular Weight; D009757:Nylons; D011506:Proteins / Q000032:analysis*; D013194:Staining and Labeling
D004396:Coloring Agents; D008567:Membranes, Artificial; D009757:Nylons; D011506:Proteins; D003101:Collodion