Most Peculiar Problem with an enzyme activity
Posted 20 January 2005 - 05:44 PM
One day I used intact cells and boom! There was activity. Lots of activity. I fractionated the whole cell batch - membrane/cell wall/CFP/cytosol/ everything. The activity disappeared once again. What is happening? where and how is the decarboxylase functioning from and why is it going away just like that?
The system is prokaryotic Mycobacterium. I need your imagination, not your expertise, so please feel free to post at will.
Posted 20 January 2005 - 07:53 PM
Mercifully free of the ravages of intelligence - Time Bandits
Posted 23 January 2005 - 05:41 PM
have you used any gfp or staining to check the localization of your enzyme? have you also used western blot to detect if there is any target in your extract? it could be that you didn't get the protein in your extract at all. if you did get your protein, then it is the question how to not let it become latent.