As I am new to the 2D gel electrophoresis technique, I have a number of questions. Basically, we perform the 2D gel electrophoresis with carrier ampholyte.
1) how to track the mobility of the protein in the IEF tube gel?
2) We used to run the IEF at
200 v for 2 hrs
500 v for 2 hrs
800 v for 16 hrs.
But in some references, it is given that once the protein reach their pI, there will not be any current flow and thus it gets stopped. Is it so?
3) I got a plain gel with a single streak. Does anyone help me in this regard?