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[Chlamydomonas] Chlamy Fluorescent Immunostaining

Heloisa Ciol via chlamy%40net.bio.net (by helociol from gmail.com)
Mon Jan 25 08:09:00 EST 2016


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


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