Two‐dimensional polyacrylamide gel electrophoresis in micro‐gels: Gradient gel electrophoresis followed by dodecyl sulfate gel electrophoresis

Abstract

A miniature system for two‐dimensional polyacrylamide gel electrophoresis is presented. Discontinuous sodium dodecyl sulfate gel electrophoresis in micro‐slab gels following high resolution micro‐electrophoresis in cylindrical continuous polyacrylamide gradient gels separates macromolecules after an initial size fractionation into their subunits. The construction of a glass cuvette for micro‐slab gels and of a suitable electrophoresis tank is described in detail. Furthermore, procedures for silver staining and fluorography initially developed for gels of normal size were adapted to micro‐slab gels. Crude tissue extracts of Funaria hygrometrica L. Sibth. were analyzed without any further purfication. Twenty samples can be processed in about eight hours.

Protein synthesis in embryonic tissues during mouse postimplantation development

“…The 10 pl samples were loaded onto the IEF gels. For the second-dimensional separation, sodium dodecyl sulphate (SDS) slab gel electrophoresis was carried out on homogeneous gels (10% w/v T, 2.6% w/v C) with stacking gels (3% w/v T, 2.6% w/v C) on top of the separating gels, as described previously [54]. More than 200 gels were processed for fluorography according to standard procedures [55,56] and exposed to preflashed x-ray films (Kodak XAR 5 ) at -8OOC [57].…”