cDNA library screening with antibody

Chris LaRosa clarosa at biocomp.unl.edu
Mon Jul 24 08:57:20 EST 2000



Warren Gallin wrote:
> 
> Tony,
>         The problem with that approach is that the cells used for immunization
> lyse and you get an immune response to intracellular proteins as well.
>         So there is no reason not to screen a library like this, your positives
> will not be biased toward membrane proteins much.
> 

One possible problem... if you used undenatured cell proteins for
antigen... the proteins on blots will be denatured... However, it might
still be useful for your purposes.  Sounds like your going to get
numerous positive clones and numerouse false positive clones with such
heterogeneous antigen and antibodies.   Be prepared for a very tiresome
and laborious screening ordeal.






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