Staking gel or spacer gel: a large pore poly acrylamide gel (4%) which is prepared with Tris buffer pH 6.8 of about 2 pH units lower than that of electrophoresis buffer. These conditions basically provide an environment for Kohlrausch reactions, as a result, proteins are concentrated to several folds and a thin starting zone of the order of 19 microns is achieved in a few minutes. This gel is cast over the resolving gel. The height of the staking gel region was always maintained more than double the height and the volume of the sample to be applied.
Resolving gel or Separating Gel: A small pore polyacryl amide gel (3 - 30%) which is prepared using Tris buffer used is of pH 8.8. In this gel, macro molecules separate according to their size. Normally in experiment, 8% gel for separate 24 – 205 kDa proteins, 10% gel for separate 14-205 kDa proteins and 12% gel for separate 14-66 kDa proteins.
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