Further improvements on the separation of alkaline phosphatase isoenzymes by gradient pore electrophoresis

Abstract

Gradient pore electrophoresis is an excellent method for the separation of alkaline phosphatase (AP) isoenzymes. There are problems associated with brittleness of the gel at high acrylamide concentrations, when 5% of total acrylamide is the crosslinking agent N, N'-methylene bisacrylamide (BIS), as is generally used. If the concentration of BIS is reduced too much the gels undergo excessive swelling on subsequent treatment. The use of a gradient gel with a constant BIS concentration of 2 g/l overcomes these problems. Gradients with this concentration of BIS also show improved separation of AP isoenzymes, with all isoenzymes so far investigated giving bands of different mobility. The use of thin layer slabs with linear gradients of 100 g/l to 200 g/l and a constant BIS concentration of 2 g/l offer an excellent method for separating AP isoenzymes. Gradient pore electrophoresis is an excellent method for the separation of alkaline phosphatase (AP) isoenzymes. There are problems associated with brittleness of the gel at high acrylamide concentrations, when 5% of total acrylamide is the crosslinking agent N, N'-methylene bisacrylamide (BIS), as is generally used. If the concentration of BIS is reduced too much the gels undergo excessive swelling on subsequent treatment. The use of a gradient gel with a constant BIS concentration of 2 g/l overcomes these problems. Gradients with this concentration of BIS also show improved separation of AP isoenzymes, with all isoenzymes so far investigated giving bands of different mobility. The use of thin layer slabs with linear gradients of 100 g/l to 200 g/l and a constant BIS concentration of 2 g/l offer an excellent method for separating AP isoenzymes.