I had my protein sample in 8M urea, and I sent my samples to another lab to perform 2DE. The people there only concentrated the sample using 10 KDa Amicon Ultra spin column, does this step consider as protein precipitation during sample preparation for 2DE? Or only what I performed in urea is consider as protein precipitationor both?
Hi Eric,
Dissolving your protein in a urea-containing solution is done to completely denature/dissolve the proteins present prior to analysis.
Protein precipitation (i.e. by addition of organic solvent) can used to clean up a protein sample, for example to get rid of urea or other (buffer) salts.
Amicon spin filters can also be used for this purpose, through a process referred to as buffer exchange; in this case however, the protein remains in solution.
So, in terms of sample clean up, buffer exchange can be considered similar to protein precipitation.
Hope this helps!
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology
- Enzymes