I've always struggled with concentrating and washing my proteins after purification. Often I lose almost everything in the process. I have used dialysis membranes followed by column filter centrifugation to concentrate or straight up column centrifugation with subsequent PBS washes.
My protein is about 50 kDa in size and I am using 10 kDa cutoff membranes/filters. When I run a gel with the elutions everything seems fine, but after washing and concentrating I have very low or even undetectable levels of protein using a microBCA assay (which is very sensitive).
What do you usually do with your eluted fractions containing imidazole in order to get a concentrated protein solution in buffer?
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