Right off the bat, I discover that my dialysis tubing leaked and my beautifully purified protein occupied a 2L volume. I first tried concentrating it, which worked at first until the apparatus broke. After that, I just loaded the remaining 1100 ml over a column.
The unfortunate part is that this protein is one in a set of five (a parent protein and four mutants) that I purify simultaneously. I prefer this approach because it ensures that the proteins are subjected to almost the same conditions. These proteins are very sensitive to purification and storage conditions, including length of time spent in storage, so it's really not ideal, in my opinion, to purify this protein by itself. So, if this protein isn't salvageable, I'll probably start over again with all five. Ugh. It's not the end of the world, but I am irritated because this dialysis was the final step and means I essentially wasted the majority of this week.
oh. well. breath in, breath out, move the fuck on.
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1 comment:
Ugh. I had a bad day experiment-wise today too, but not quite so bad as yours sounds. Hope it turns around with the next go at it.
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