We have a huge problem with a second dimension (SDS-Page) during 2-D
electrophoresis protocol. After isoelectric focusing, during SDS-Page, proteins leave stacking ("upper") gel, move fluently to analytic
("lower") gel and then do not move to lower part of analytic gel. When proteins should move from upper part to
lower one, they are "blocked" and after that, only pigment
(bromophenol blue) moves down. We've changed all reagents, pH of
buffers is proper. We've tried to use three different ACs and three
different equipments for electrophoresis. Electrode buffer is prepared
properly too. Second dimension electrophoresis runs on constant
intensity, but we've observed that volts are rising very fast.
Of course, it is not our first attempt. Before we don't have any
problems like that.
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2 D electrophoresis _ proteins do not move to lower part of analytic gelSDS- PAGE 2D electrophoresis
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