1) In pursuit of the goal of a 2-dimensionally steady-state macromolecular map, the filed strength across a pH 4-10 gradient was made more even by the addition of carrier ampholytes to Immobiline gets. This eliminates a) lateral zone spreading; b) the pile-up of protein zones at the edge of low current regions around neutrality, c) the retnetion of proteins in the sample zone, and d) allows for pH and voltage measurements not or difficulty feasible without such addition. 2) To obviate the considerable gel length associated with flat pH gradients over the wide pH range (4-10) and the resulting need for very high total voltage across such gels, a diagonal mode of Immobiline electrofocusing was devised. 3) The effective radius of the agarose fiber was determined in the order of 1 nm, using high agarose concentrations suitable for the sieving of proteins. This shows that the reduction in fiber radius from about 18 nm, found with viruses, to the same value previously determined for hydroxyethyl-agarose is not due to hydroxyethylation. 4) The divergent effective fiber radii of agarose found with viruses and proteins were reconciled by the demonstration of non-linear Ferguson plots for both, and thus increasing KR, decreasing r, increasing l' and decreasing fiber volume (ml/g) with increasing agarose gel concentration. 5) Ferguson plots were derived for electrophoresis in linear, uncrosslinked, liquid polyacrylamide. Sieving is qualitatively similar to that obtained with crosslinked gels. 6) The potential application of the capillary apparatus with optics for sieving of macromolecules with liquid polyacrylamide was initiated. 7) A device for rapid diffusion of gels was constructed by use of which the staining time of SDS-gels can be reduced bo 1.5 h or less. A similar device for the silver staining of cylindrical gels is being developed.
Showing the most recent 10 out of 20 publications
