in-situ protocol
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From: | DDittus787@aol.com |
To: | histonet@pathology.swmed.edu |
Reply-To: | |
Content-Type: | text/plain; charset=US-ASCII |
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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