Hi Greg,

I've had limited success with TNF-alpha (Pharmingen) 1:100 on mouse footpad infected with Leishmania major. This study was a time course and I only saw it late in infection. That being said I used Formalin fixed tissue 24 hrs , paraffin embedded,  a ABC protocol, a biotinylated Rat secondary (vector Labs), and Trypsin digestion at 37 degrees C for 20 minutes. The only reason I believe its is real is that the other time points where negative and the isotype control was negative. It's my though that you need a lot of cytokine around to pick it up my immuno.   That's my limited success story hope it helps. If you need more details let me know.

Jamie Erickson 
Genetics Institute



>>> Greg Dobbin <dobbin@Upei.CA> 08/19 11:38 AM >>>
Hi All,

Can anyone tell me their preferred method of Ag retreival for Tumour 
Necrosis factor in mouse tissue (Formalin/paraffin)? Heat or steam? 
Which buffer or enzyme?  Thanks. Greg
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Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 Unviversity Ave.
Charlottetown, P.E.I.
C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851

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