I have a protein known to bind to nucleic acids. I expressed it, lysed the cells using a microfluidizer and spun down the lysate. I then collected the pellet and resuspended it in a buffer containing 8 M urea.
Hereon the solution turned into a slightly viscous fluid (bubbles suspended in the solution rise moderately slowly as opposed to instantaneously in water). I spun this solution down again at 50000 g. The viscosity remained.
I wish to load this onto a nickel column but would like to filter the solution first. The problem is that both 0.2 micron and 0.45 micron filters get clogged and filtration seems impossible.
Could you suggest a way to make the solution more fluid so that it can be filtered? I would prefer not to dilute the solution too much.
I imagine the protein and nucleic acids are causing some kind of polymeric network inside the solution? Could you also suggest a way to remove the nucleic acids?
Increase the ionic strength of the solution (e.g. by adding concentrated NaCl or dissolving the reagent right into it) to weaken the ionic interactions between the protein and the nucleic acids. You can also try treating the solution with nucleases.
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