Fix for zfish & GFP?
chien at mpib-tuebingen.mpg.de
Fri Feb 21 11:40:32 EST 1997
Here's an expanded version of the second response that I just sent to Yiing:
Yup, it looks dimmer after fixation, all right. Why don't you try fixing
overnight at 4 degrees and see how this works? I also take it that you're
making up 4% PF fresh from powder.
I don't expect the S65T to affect fixation characteristics; after all, the
chromophore is still buried deep inside the protein. I have to admit that I
haven't done so many before-and-after comparisons, but I really wouldn't
expect the GFP to fade so much. The question of fixation conditions was
discussed several months ago on the GFP newsgroup, and the gist was that
insufficient fixation can be a problem. I believe that methanol may quench
the fluorescence, but you're not using it.
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