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I use both protocols (and normally I dont check amount of protein, as I use the same amount of cells and I am not working with cell death or apoptosis or something that kill my cells).
If I want to see some specific protein modification (as sumoylation) I use laemli directly on the samples, but to avoid the viscosity that is how I do it:
Recolect the cells
Add laemli buffer 1x 120ul
Boil for 2 minutes
Sonicate 2x15sec
Boil 5 minutes
I have no problem with this easy protocol
#37655 direct cell lysate
Posted
on 25 September 2009 - 04:34 AM
