High Acrylamide Concentrations Research Articles

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

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.

Separation of the messenger RNAs of Newcastle disease virus by gel electrophoresis.

We have separated the 18-22S putative messenger RNA of Newcastle disease virus into seven species ranging in molecular weight from 0.55 to 1.53 x 10(6) using sodium dodecyl sulfate-acrylamide-gel electrophoresis at relatively high concentrations of acrylamide and for a relatively long time. Studies of the number and molecular weights of the proteins and the 18-22S RNAs of the virus suggests that these RNAs are in the right molecular weight range to code for the known proteins of Newcastle disease virus. In preliminary studies using this separation technique, we have demonstrated that: (a) there is no difference between the 18-22S RNA made during a normal infection and when genome replication is blocked; and (b) there is a strain-specific difference between the RNAs of Newcastle disease virus-AV and Newcastle disease virus-HP.