Simultaneous immunoblotting analysis with activity gel electrophoresis and 2-D gel electrophoresis.
Diffusion blotting method can couple immunoblotting analysis with another biochemical technique in a single polyacrylamide gel, however, with lower transfer efficiency as compared with the conventional electroblotting method. Thus, with diffusion blotting, a protein blot can be obtained from a sodium dodecyl sulfate polyacrylamide gel for zymography assay, from a native polyacrylamide gel for electrophoretic mobility shift assay (EMSA), or from a two-dimensional (2-D) polyacrylamide gel for large-scale screening and identification of a protein marker. Therefore, a particular signal in zymography, EMSA, and 2-D gel can be confirmed or identified by simultaneous immunoblotting analysis with a corresponding antiserum. These advantages make diffusion blotting desirable when partial loss of transfer efficiency can be tolerated or can be compensated by a more sensitive immunodetection reaction using enhanced chemiluminescence substrates.
Graduate Institute of Biochemical Sciences, National Taiwan University, PO Box 23-106, Taipei, 106, Taiwan.
Der-Yen
DY
Lee
Geen-Dong
GD
Chang
D000818:Animals; D002347:Carps / Q000033:anatomy & histology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods; D015151:Immunoblotting / Q000295:instrumentation / Q000379:methods; D051379:Mice; D008862:Microsomes, Liver / Q000737:chemistry; D011506:Proteins / Q000032:analysis*
D011506:Proteins