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Too much precipitate. Help out a first timer!

Too much precipitate. Help out a first timer! - Protein Crystallography Forum

Too much precipitate. Help out a first timer! - Discuss research on protein crystallography and crystalization. Protein Crystallography Forum.


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Old 11-04-2011, 02:56 AM
Pipette Filler
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Default 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
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Old 08-24-2012, 04:33 PM
Pipette Filler
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Default Re: Too much precipitate. Help out a first timer!

check the concentration of your protein.
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