Dear all,
I was wondering if anyone could share here some experience in Chlamy
fluorescent immunostaining, please?
Since I started in this field, I couldn't apply to my experiments any of
the protocols mostly used so far. For some reason, I cannot fix my cells in
coverslips (I prepared coverslips at the lab, either using poly-L-lysine or
polyethyleneimine). At first the cells seem to be stuck at the glass, but
once I go to the washing steps, almost all of them disappear, and in the
end, I can find 2- 5 cells at the whole coverslip. To overcome that, we
developed a protocol for cell incubation in eppendorfs with blocking and
antibody solutions, which gives us a lot of cells, but not an uniform
staining pattern.
Besides that, we are trying to use alpha-tubulin or
alpha-acetylated-tubulin commercial antibodies as well, but we couldn't
stain a single cell so far.
Does anyone have a tip or a protocol to share that I could try using here
in our lab?
Thank you all.
Best regards,
--
Heloisa Ciol
