Dot Blot with plasma membrane Proteins

Low Teck Yew (medp9148@nus.edu.sg)
Thu, 18 Nov 1999 07:48:12 +0800

Hi all,

I am doing a dot blot with a plasma membrane protein which I had purified
without adding in any solubilization agent. This is followed by
immunodetection with polyclonal antibodies. The results I got is negative
and the nitrocellulose membrane shows negative staining. I am not too sure
if the antibodies do not bind to the membrane proteins or I should
solubilize the membrane proteins first with a non-ionic detergent like
Triton-X before I do the dot blot? If this is the case, what is the best
way?

Thanks!