rat intracellular TNF-alpha

From: Julie_Wieseler-Frank/CAS/UNO/UNEBR@unomail.unomaha.edu
Date: Thu Oct 12 2000 - 08:06:24 EST 

Hi!

I am trying to measure intracellular TNF-alpha in rat splenocytes -- the
operative word is trying.  I am stimulating with LPS -- I have tried stimulation
times of 30 min to 13 hours; I have used monensin and brefeldin; i have used
plastic flasks (scraped prior to staining cells) and I have tried siliconized
tubes.  When I look at cultures under the microscope, the morphology of the
cells indicates activation.

Any insights? I would appreciate ANYTHING!!

Thanks for your time--

Julie

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