Recipe needed for native gel - (Mar/28/2007 )
I needto run Native gel for my protein to esimate the aggregation size.PI of protein is 4.6. Can anybody recommend me what should be ph of the resolving, stacking gel and running buffer. what about the composition of the different buffer ? I used to run 12 % SDS-PAGE.
You should be fine using your usual recipe and just omitting the SDS. At the standard run (pH 8.8) and stack (pH 6.5) conditions, your protein should have an overall negative charge and migrate toward the cathode.