Summary
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.
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Acknowledgements
This work was supported by grants (to G.D.C.) from the National Science Council, Republic of China.
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Lee, DY., Chang, GD. (2009). Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel Electrophoresis. In: Kurien, B., Scofield, R. (eds) Protein Blotting and Detection. Methods in Molecular Biology, vol 536. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-542-8_4
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DOI: https://doi.org/10.1007/978-1-59745-542-8_4
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