[Histonet] Re: IHC on coverslips

From:"Sarka Lhotak"

Hi Netters,
IHC on coverslips is easily done by performing the whole procedure,
including fixation and permeabilization with Triton X, in a 6 well
plate. Label the lid of the plate to know which slip is in which well.
Rinse by filling the well with buffer, then aspirate. Before
incubations, I aspirate as much liquid as possible from around the
coverslip with a glass Pasteur pipette. This will limit spreading of
your precious antibody into the well. Then I carefully place 5 drops of
the antibody (50 ul), in the middle and four corners of the coverslip. 
I use this technique mostly to do immunofluorescence, but I have also
done peroxidase/ AEC reactions, with great results. The coverslip is
removed at the very end for "coverslipping" with a slide. The coverslip
is cells up all the way, just make sure you coverslip it cells down.
Hope this helps. Please, contact me for more details.

Sarka Lhotak, PhD

McMaster University
Hamilton, Ontario, Canada

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