I am facing problem while running SDS PAGE. Stacking gel is shrinking and the wells are not forming properly. Bands are also not coming despite the enzyme is showing activity........Can anybody help me in sample preperation and appropriate percentage of gel for SDS PAGE for the protein having MW between 60Kd- 95KD
Thanks

to prepare 10ml of stacking gel, I use:
6,8ml H2O
1,7ml 30% acrylamid mix
1,25ml 1M Tris (pH 6,8)
0,1ml SDS 10%
and just before use
0,01ml(/ml of gel needed) 10% ammonium persulfate and 0,001ml (idem) TEMED

It works well

be sure to drain out the water completely after resolving gel is poymerise and also do cast 10%gel for ur purpose
Do quantify the protein before u load onto gel

vaishali said:

I am facing problem while running SDS PAGE. Stacking gel is shrinking and the wells are not forming properly. Bands are also not coming despite the enzyme is showing activity........Can anybody help me in sample preperation and appropriate percentage of gel for SDS PAGE for the protein having MW between 60Kd- 95KD Thanks

Sometimes if the APS is not fresh, then the gel can't form properly. Make sure to use good APS.