Hi,
Can someone please suggest which detergents to use in regular lysis buffer/resuspension buffer for E. coli cells? Please also suggest the optimum concentration to be used.
Thanks.
It depends on what you wish to accomplish. Do you want to solubilize everything in the cells but don't care whether the proteins are in the native state? Do you want to dissolve membrane proteins and keep them in their native state? Do you want to lyse the cells, keep soluble proteins in the native state, but don't care about the membrane proteins?
As Dr. Shapiro suggested it depends on the application and compound of interest. DNA/RNA/Proteins/Lipids all have slight difference in protocol in terms of cell lysis.
@ Adam: Can you plese suggest the detergent if I want to keep soluble protein in native state and also detergent if we want to keep membrane protein in native state?
Is there any mild detergent that can keep both soluble and membrane proteins in native state but only help in cell lysis??
You should use a non-ionic or zwitterionic detergent. Triton X-100, Zwittergent 3-14, n-octylglucoside, CHAPS, and dodecylmaltoside are some examples. You will have to experiment with the detergent concentration needed to dissolve the membranes. It should exceed the critical micellar concentration of the detergent, and it will depend on the cell concentration.
