Use of MLH1, MSH2 and MSH6 antibodies + preparation of airdried s mears for IHC.

From:Sherna Coote

Hi 

Does anyone have any experience of using MLH1, MSH2 and MSH6 antibodies.  We
are currently testing these antibodies and I would appreciate some input.
We are currently, blocking in peroxidase block of Dako En Vision kit for 7
minutes, microwaving for 20 minutes in citrate buffer (pH 6), incubating in
Ab for 2 hours at room temp, incubating in Dako Envision Polymer for 2
hours, then visualising using DAB.  As you can see this method takes up most
of  the day and I would like to cut down the incubation times if possible.

Also, how do people prepare airdried smears for immuno.  Not had much
experience of this but my current lab fixes in Van der Griendt's fluid (
0.1% formal saline) for 2 hours!!

Thanks in advance.

Sherna Coote.
Grade Technologist.



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