Separation and radioautography of microgram quantities of ribosomal proteins by two-dimensional polyacrylamide gel electrophoresis

摘要: Recently a two dimensional polyacrylamide slab gel electrophoresis system was described by which all of the ribosomal proteins E. coli were separated into distinct spots [ 11. This and others have been used to separate eukaryotic as well prokaryotic [2-61. The developed KaltSchmidt Wittmann [l] gives excellent resolution both types proteins; moreover, standard highly useful nomenclature ribosoma1 has defined in terms this ]71. We adopted basic Kaltschmidt use thinner smaller slabs require amounts sample, can be dried for radioautography radioactive proteins. advantages two-dimensional here are summarized follows: Ribosomal protein required: 50 100 pg; total time: 12 hr; staining destaining 3-5 radioautography: readily possible; totally 2-3 hr without distortion or shrinkage accurate radioautograms; estimated cost apparatus, about $ 160 (not yet commercially available). with miniature is essentially identical that reported It also employed identify those phosphorylated kinase [8].