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-   -   Too much precipitate. Help out a first timer! (http://www.molecularstation.com/forum/protein-crystallography-forum/83956-too-much-precipitate-help-out-first-timer.html)

BeastfromthEast 11-04-2011 02:56 AM

Too much precipitate. Help out a first timer!
 
Hi,
I'm trying to crystalliza a protein inhibitor complex. I added 1 microliter of 100 mm NADP, 1 microliter of 100 mm DUMP, and 1 microliter of 50 mm inhibitor to 50 microliters of protein. of incubated them. In the wells I put a mix of Sodium Phosphate, Potassium, Ethylene Glycol, Phosphate Citrate, and Water. The hanging drop was 2 microliters of the protein, 1 microliter of water, and 1 microliter of what was in the well.

I cant list all the combos of chemicals in the wells, but I'll list a few:

55 microliters sodium phosphate, 55 microliters potassium, 22.5 microliters ethylene glycol, 50 microliters phosphate citrate, and 67.5 microliters of water in the first well. The other wells were pretty similar to that.

I got a lot of precipiate when I did this last time and my PI wants me to figure out what is the problem. Thnaks

vakkomak 08-24-2012 04:33 PM

Re: Too much precipitate. Help out a first timer!
 
check the concentration of your protein.


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