Western Using Monoclonals

[Annette C. Hollmann]

In article <Pine.3.89.9410302005.C95666-0100000 at blue.weeg.uiowa.edu> qiwang at BLUE.WEEG.UIOWA.EDU (wang qi) writes:
>
>Dear netters,
>I am have some trouble with  western blotting using monoclonal 
>antibodies.
>Several monoclonal antibodies have been generated in our lab by injecting 
>mice with whole cells. One of our antibodies recognizes an antigen on the 
>cell surface using indirect-immunoflouresence microscopy.  
<snip>
>We, however, have not been able to identify any specific band 
>on the immunoblots.

Your monoclonal may recognize a conformational epitope that is destroyed
by SDS-PAGE. If you have a different monoclonal that reacts with the same
protein,  try it. It just might work.
I have done Western Blots with both polyclonals and monoclonals, and have
found that some monoclonals work beautifully while others don't work at all.
Also, some monoclonals that can be used for immunoprecipitation can not be
used for Western blotting.

Annette
ah690549 at mbcr.bcm.tmc.edu