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Cell Lysis for Enzyme Activity

Cell Lysis for Enzyme Activity - Cell Biology and Cell Culture

Cell Lysis for Enzyme Activity - Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology.


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  #1  
Old 08-27-2007, 12:12 PM
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Question Cell Lysis for Enzyme Activity



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
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Old 07-19-2009, 12:53 PM
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Default Re: Cell Lysis for Enzyme Activity

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!

Last edited by ConsultEnzy; 07-21-2009 at 05:28 PM.
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Old 02-22-2012, 08:05 PM
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Default Re: Cell Lysis for Enzyme Activity

thanks
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Old 03-05-2012, 04:58 AM
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Default Re: Cell Lysis for Enzyme Activity

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

Thanks for the help.
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Old 11-08-2012, 01:40 PM
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Default Re: Cell Lysis for Enzyme Activity

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
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