Methanol Fixation

Andrew Doherty A.Doherty at Bris.ac.uk
Mon Oct 28 12:25:11 EST 1996


Hi everybody

A nice simple question - if I fix some cells using methanol at -20 deg
C, do I lose all the soluble protein, just leaving the membrane bound
protein behibd to be detected? The reason I ask, is that cells I have
transfected with GFP (soluble) do not fluoresce after fixing in this
way, whereas those transfected with a receptor-GFP fusion protein do
show fluorescence. I also know that in live cells I get very good
fluorescence with the GFP alone. So does this mean that I'm losing the
soluble GFP on fixation, but retaining the GFP when it's fused to a
membrane bound receptor?

Ta in advance

Andy D

email A.doherty at bris.ac.uk



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