[Histonet] Roche TUNEL kit

[Margaret Blount]

Dear Katy,

Thanks for sharing your expertise with us.

Margaret
Margaret Blount
Chief Technician
Clinical Biochemistry
University of Cambridge
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2QR 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Katy
Whalley
Sent: Thursday, June 03, 2004 9:16 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Roche TUNEL kit


Hi,

To everyone who asked for the TUNEL protocol, here is the one I use, which
seems to work well. Give it a test run, and I hope it helps,

Katy

In Situ Cell Death Detection Kit (Cat no. 1684795)

1. Dewax and Rehydrate sections as usual

2. Permeabilise tissue -- incubate with following Proteinase K Sol for 15 
mins at room temp:
   for 1 ml:       915ul dH20
                   50ul Tris (1M)
                   25ul EDTA pH 7.4(200mM)
                   10ul Proteinase K (1mg/ml)
   Solution must be pre-incubated for 30 mins at 37 degrees before use

3. Rinse sections in PBS 3x5 mins

4. Prepare TUNEL reraction mixture ON ICE immediately before use:
                  5ul enzyme solution: 25ul labelling solution per section

   Incubate sections with TUNEL reaction mix for 1 hour at 37 degrees

5. Rinse PBS 3x5 mins

6. Mount and coverslip


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