These are Maltose Binding Protein crystals and I have to open the seal to make a buffer transfer. The moment I open the wells, within 2 mins the solution turns opaque as shown. Can someone help me what exactly is happening in these solution? How can I avoid this ? Anyone else faces this problem?
Dear Amar,
It is extremely difficult to say accurately what is happening in your solutions without knowledge of exact crystallization conditions and the compositions of the buffers. One only can speculate that change in oxygen content, pH change, partial pressure change of volatile agents change etc. is the cause of the visible change which is an obvious precipitation or denaturation of an agent. Whether this is the protein or solute it is difficult to say. I would bet that this is a protein component.
Two pieces of advice.
(1) Why would you disregard the precipitate and work with what you have.
(2) If you really worry about the change why you do not limit the change by working in a cold room, or in the room with humidity close to the well, or alternatively use a portable plastic manipulation bag with water or alcohol or any other agent in it.
You can use a regular microscope to do you manipulations.
My first approach would be to try to freeze the crystals directly from the well in the gas stream and forget about any transfers. If you need to soak it then you need to worry about my advise.
Good Luck.
Bog
Likely this happens because of evaporation of water from the (very small) drop. Depending on the composition of the buffer/precipitant, one of them may precipitate - it could even be your protein. What I usually do if I need to work with the crystals is to carefully add a bit of stabilizing solution to the drop. A stabilizing solution should contain the components of your reservoir solution, with the precipitant concentration a little higher than that of the reservoir (sometimes also the components of your protein solution should be added, but you can first try without those).
Depending on what you want to do with the crystals, they can also be transferred to a larger volume (say 500 ul), this will eliminate the problem of evaporation altogether.
Hope this helps, good luck !
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