Isozyme electrophoresis

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AnjaliKrish
AnjaliKrish's picture
Isozyme electrophoresis

HI,
  I'm working with isozyme electrophoresis for population variation study I'm getting only  single band for all enzymes like ADH,LDH etc and i'm using 6% resolving gel 5% stack gel I tried 7%,10% gel but i got diffused bands and it takes more than 3 and a half hours.I'm running out of my time to submit my thesis only 1 week more Please help me

Chin Fen Teo
Chin Fen Teo's picture
Hi AnjaliKrsih,

Hi AnjaliKrsih,

It depends on how big difference are there between all these isozymes. If they have indeed very similar in size, 1D-PAGE with regular running condition is almost impossible to separate them. While increasing the electrophoresis time, you are indeed also increase the resolving power of molecules/proteins with very similar size. I suspect, letting the gels run extra-long might be the only way to go for 1D-PAGE separation.

Good luck.