Greetings: Regards Steve Kayes Dr. Stephen G. Kayes, Professor Structural and Cellular Biology; 2042 MSB University of South Alabama Col. of Medicine Mobile, Al 36688-0002 (334) 460-6768 (Ofc) (334) 450-6771 (Fax) >>> Tina Rogers <Tina.Rogers@ccc.uab.edu> 19 October 1999 4:35 pm >>> Greetings. I have a user that needs to isolate granulocytes on a large scale from mice. Does anyone have experience or suggestions? Thanks in advance. TIna S. Rogers, Ph.D. Manager, MAMDC FACS Core Facility Division of RheumatologI have successfully used a 10% solution of proteose-peptone (sterilzed) injected ip (1.5 ml) per mouse. Between 6 and 12 hours later you should be able to isolate peritoneal exudate cells (PEC) which are almost pure neutrophils. If you let the mice go longer before performing peritoneal lavage, you will begin to get macrophages and other resident cells and the neutrophils will be mostly disintegrated. BTW, if the mouse happens to harbor a tissue dwelling helminth infection, you can lavage at 48 hr and get almost pure populations of eosinophils by this same technique. y University of Alabama at Birmingham 1900 University Blvd. THT 405 Birmingham, AL 35294-0006 205-934-1362
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