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what do I need to perform protein crystallization?


Justin.Frank

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I want to make my protein crystallize for electron microscope to study their structure.

 

For what purpose? For the purpose of identical replication or to study their differences under varied conditions?

 

Protein crystals are highly irregular structures and environmentally sensitive. Likewise, there are several thousand conditions unique to each protein, so it's best to use a known protocol for the material in use.

 

I'm not sure what your objectives are at this time, but can suggest a couple of things that may be helpful in your research.

 

Micro batching is the simplest method by placing a supersaturated protein sample within an inert oil medium and left to stand undisturbed, but the resulting crystals will undoubtedly vary in structure and will not handle well without changes.

 

You might try vapor diffusion instead. In this method, a drop of protein is treated with a buffer to maintain pH and a precipitant (polyethylene glycol) then placed in an open vessel within a sealed vessel containing a similar solution, but in higher concentrations. Allowing the two to equilibrate may provide an opportunity to observe crystalline structures once formed. Unless you do this in multiple samples and view them incrementally, it's difficult to observe these changes as they occur. Unless of course, degradation of these crystals is something you're interested in.

 

There are other methods. Such as dialysis, but that's out of my league.

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Protein crystallization has always been part alchemy. Finding the right condition is always a bit hit and miss. With an unlimited budget automated platforms are ideal as they can run through a lot of conditions. There are also dozens of techniques that let you move through conditions rapidly. Hanging drops are probably the fastest screening method if done by hand and also is amenable for automated observation.

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