why it is not showing zone?
for how long the protein remain active after , precipitation, if kept in buffer?
HI Jannatul,
Please provide more information about your question.
Yes, I agree with Dyoni. You need to clarify your question for getting relevant answers, comments or suggestions for your problems. Any ways, proteins are fragile molecules. They may be degraded or converted into their unstable forms at any stage during upstream or downstream processing or upon storage. If your protein from the crude cell lysate shows activity, then you need to check the condition of your precipitation step. What was the conditions of your precipitation? what do you mean by 80% precipitation? Did you use salting-out precipitation using ammonium sulphate or sodium sulphate or other salts? or Did you use organic solvents, such as acetone or ethanol? That is why we are saying the information you have provided is not sufficient to give relevant answers. I guess your precipitation condition might have affected the stability of your protein. Thus you need to:
1. optimize the precipitation conditions, such as concentration of salts, organic solvents, temperature, time or speed of agitation (if necessary) and conditions of the buffer for re-suspending the protein pellet after precipitation
2. consider other purification techniques, such as filtration, chromatographic-based techniques.
I hope these helps. Good luck!
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