Hi every one,
I am doing my research on Helicobacter pylori. After lyse the infected mammalian cell, I am doing western blotting to check the injected virulent protein. for that I am using rabbit polyclonal abs, and am experiencing non specific binding and lot of noise background, mean while i didnt experience any other non specific binding while using monoclonal abs for other targets.
Am doing 1:30 hrs membrane blocking, abs incubation for over night, 1:30 hrs abs blocking and 1:30 hrs secondary ab incubation with 3x10 min washing. some times i didnt got any noise but many times i am getting noise background and non specific binding.
How to overcome this problem.
Hope for looking valuable suggestions.
micronaguMember Since 21 Aug 2012
Offline Last Active Mar 14 2013 06:50 PM
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My research interests
Infection biology, Microbiology, cancer biology
30 Aug 2012 - 23:35