Western Using Monoclonals

[Mic Chaudoir]

QiWang,

The previous posters are right.

1) Membrane proteins are often differentially glycosolated, which could
generate the smear you are seeing.  You can treat your samples with
N-Glycanase, if you want, to look at this problem.  N-Glycanase will
remove the sugar moeities.  

2)  Your protein may be denatured on the gel, and the epitope destroyed. 
Try using a polyclonal (mouse serume before the animal was sacked to make
hybrids).  Failing that, you can try running a non-denaturing gel.

Good luck!

-- 
mic
mic at nwu.edu

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