in-situ protocol

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From:DDittus787@aol.com
To:histonet@pathology.swmed.edu
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Dear All
After many crazy days, being out of town, and reviewing my old mail, i 
discovered that some asked for my  in-situ protocol, so here goes, please 
accept my apologies.
oven set to 105 degrees c
deparraffinize slides
wash slides with di h20
pepsin for 3 min at 105
probe for  5 min at 105
remove slides
oven at 45 degrees-place slides in oven when oven reaches 75
leave in oven as it reduces to 45-incubate here for 45 min
wash slides in 2x ssc 2x at rm temp
incubate with strep-avidin-hrp for 5 min at50degrees c
wash with probe lock
incubate with dab 5 min room temp
counterstain with hematoxylin and coverslip
The above procedure was developed by our director- Dr Kathleen Montone on the 
microprobe system from cms-fischer , but works nicely by hand
We use reagents from Research genetics-great stable DAB.

Sorry for the time delay---------------------Dana



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