Percentage of SDS- PAGE gel for co-IP - (Aug/22/2006 )
Dear friends,
I am doing co immunoprecipitation for yeast two hybrid system. The problem is I don't know how I should calculate the percentage of SDS-PAGE gel. I mean should I add the size of two proteins together and based on that prepare the gel or I just have to consider the size of protein which I use primary Ab against that?
HELP please
-science-
QUOTE (science @ Aug 22 2006, 08:27 PM)
Dear friends,
I am doing co immunoprecipitation for yeast two hybrid system. The problem is I don't know how I should calculate the percentage of SDS-PAGE gel. I mean should I add the size of two proteins together and based on that prepare the gel or I just have to consider the size of protein which I use primary Ab against that?
HELP please
I am doing co immunoprecipitation for yeast two hybrid system. The problem is I don't know how I should calculate the percentage of SDS-PAGE gel. I mean should I add the size of two proteins together and based on that prepare the gel or I just have to consider the size of protein which I use primary Ab against that?
HELP please
When you do a coIP, you use non denaturing conditions. When you run an SDS-PAGE, since you are using SDS, you will "destroy" the interaction between the two proteins, therefore you will have them running separate.
The answer to your question is: you should run a SDS-PAGE with the right acrylamide percentage for both your proteins
-dnafactory-