I am using 2 million cells, adding equal volume of 2x sample buffer, boiling for 15 min at 95 degree, centrifuge and loading on gel.
Please suggest me What are the changes I can do in my protocol?
You need to reduce DNA viscosity either enzymatically or mechanically
It all depends what do you want to follow ...What is the MW of your target protein (s)? I think you can undergo some dialysiswith an appropiate cutoff...Do you intent to use antibodies to detect the target proteins? These answers must lead to certain choices on how to perform the best suited sample preparation protococ....
maybe you could give a little more information of what you are doing with what kind of cells and what problems you encounter.
I am doing western blotting and using DT40 chicken B cells. But everytime the bands in SDS PAGE are not looking intact.
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