Cell Lysis for Enzyme Activity

[oBWhat]

Hello all,
I am back with a question some expert may know.

I want a lysis method that does not denature the proteins much.

I know there are french presses for lysis using pressure etc. but what is really the best way to lyze cells for analysis later of enzyme activity?

this requires non-denaturing conditions and minimal disruption of the protein secondary and tertiary structure.

Any ideas?

cheers
ObWhat

[ConsultEnzy]

I use BugBuster for such applications:

[Only registered users see links. ]

It costs quite a bit on a large scale, but for small scale applications its great stuff (it will not disappoint, I can guarantee it - used it on lots of different cells). All you do is squirt it in to your resuspended cells (or even just the culture if you have deep enough pockets). Generally I compare BugBusted cell extracts to one those prepared by normal sonication. When I see no difference, then I know I can sonicate away without being worried about doing damage!

Good-luck!

[wr_1a]

thanks

[Babak]

Does anyone knows the chemical lysis agent which lyse Staphylococcus aureus and does not inhibit PCR?

Thanks for the help.

[jcoalsonshore]

This tissue pulverizer gadget works in liquid nitrogen (good for enz activity if they are delicate) and comes with a drill bit accessory to nuke tough samples. The company is called Cellcrusher, they have a video that explains how it works, google them