RE: Pig WBC

From: Mara Rocchi (M.ROCCHI@CGNET.COM)
Date: Tue Oct 21 1997 - 14:10:00 EST


Dear Dr. Cabrera,

I haven't flowed pig WBC, but cow, cat, sheep, and hamster. If you don't 
want to Ficoll your cells (dilute your blood 1:2 or 1:3), you can try to use 
an NH4Cl-based lysing solution.
An easy to prepare is: 8.99 g/L NH4Cl, 1g/L KHCO3, 37 mg/L tetrasodium EDTA, 
pH 7,3. Add 7 ml of lysing solution to 500 microliter of your blood, mix 
gently and when RBC will are lysed spin for 7 minutes. Wash 2x in saline or 
PBS and resuspend in your staining media. I usually stain 10 samples 
starting from 500 microliters of blood  (cow PMN phenotyping).

Hope this help

Mara

 ----------
Mara Rocchi, DVM, Ph.D.
International Livestock Research Institute
Nairobi, Kenya
Fax: 00254-2-631499
e-mail: M.Rocchi@cgnet.com



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