Thank you.
Protein buffer: 200mM NaCl, 50mm Tris pH:7,6 and 10% glycerol
I could not understand your question. Could you please give some more details?
Hi Aditya. Yes of course. I want stabilize the monomeric protein form ( the single disc) But I don't know how to do it. At the moment I could see to EM 80Kv the protein monomer stacked forming a row of these disc and I only want the monomeric form ( the single disc). I'm looking for some advice to stabilize such monomer
What is Mw of your protein, and what is size distribution of the stacks? How do you want to solve the structure? If by EM I would start looking at the protein as is, may be the stack has a regular shape and that's the structure you want to determine.
If by crystallography you may want to have the monomer, although the fact that the protein forms stacks could be of use when trying crystallizing it.
It all depends on what kind of interactions stabilize the stack: ionic, then increase salt; hydrophobic, try with detergents; low concentration of kaotropic agents may also help. Without knowing anything else you will have to test several conditions and see what works.
The Mw is 134.000Da 1 unit and the stack hasa Mw:1,300,000 Da. We dont care about the method to solve the structure. At the moment We are trying EM and crystallography .
The only thing I would add to Carlo's excellent answer is that you need to take care that the method you use to produce crystals does not reverse whatever you need to do to to dissociate the stacks. E.g. ammonium sulphate is very commonly used in crystallisation trials, but it is a potent agent for strengthening hydrophobic association.
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