HOW TO BUILD A SANDWICH ELISA SYSTEM? - (Sep/05/2001 )

Hi there,

I suggest that you:1-coat with a capture Ab2-add ur Ag3-add a biotinylated Ab4-develop with Strep-HRP

I WANT TO BUILD A SANDWICH ELISA SYSTEM,BUT LITTLE EXPERIENCE ABOUT IT. AND IN MY EXPERIMENT I FOUND THE VALUE OF THE OD.INCREASED WITH THE DECREASING OF THE ANTIGEN. IF ANYONE KNOW IT ,PLEASE TELL ME. THANKS.

hai wallan, you can do like this:1]coat your primary anti body keep it at 37c for 3 hours .2]come up with your antigen at 37 c for 1 hr after washing and blocking for i hr3}come up with antibody which binds to your antigen at a different epitope4] then come up with your secondary antibody this way you can reduce your background noise