hi, does anyone know what happens when a column dries out? i think there's a tear in the bottom of my column, and even though i stopped it up at both ends, it dried out. it's a his-tagged protein purification elution, and i'm just wondering if i;ve lost all my protein grrr.. any suggestions as to what i should do? i'm still going thru with my elution...

It depends on the properties of your protein. Typically, drying out a column isn't a big deal for the column unless it's something that's got to go through a specific packing protocol (like gel filtration or higher resolution Q columns). But for the protein, it will get at least partially dehydrated. If your protein happens to be particularly stable, it'll just redissolve when you elute it and you shouldn't have any problems. If it's not so stable, it will likely elute but in a denatured form.

I would analyze not only the protein content of your eluent, but also the protein content of a sample of your spun down eluent (to remove aggregates), and also the activity if you have a way to measure it.

thanks. i don't know exactly how stable my protein is since this is the first time we tried to purify it, and there's no assay to test specific activity, either. will try to work with what i have