GFP antibody staining

Sascha Rvhrig roehrig at lmb.uni-muenchen.de
Tue Aug 18 12:53:51 EST 1998


Hello, maybe somebody can help me out here:

I have worms expressing the ClonTech Enhanced GFP. They give very good
fluorescence in the living animal.. Yet, after fixing the worms for
antibody staining, I was not able to detect a single fluorescent cell
anymore. Has anybody experienced similar things when fixing worms. This
problem did not arise, when I fixed worms expressing either wildtype or
the S65T GFP variant.

Trying to stain the worms with a monoclonal GFP antibody sold by
Quantum, I ran into another problem. The antibody stains every nucleus
in the worm, even metaphase chromosomes.

If anybody has any suggestions or comments, I would greatly appreciate
it.





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