I think this link will provide you with the best answer:
http://www.jove.com/video/52749/isolation-murine-peritoneal-macrophages-to-carry-out-gene-expression
I almost did as Fernando said - I let them sit over night in simple medium and treated with LPS. Induction was beautiful. But I guess you could even do it earlier, as soon as they sit - as it's done with other tissue macrophages ... although I did not elicit them with thioglycolate, but just took naive mice - I got like 2x10^6 macrophages per mouse ...
Isolate count plate in RPMI 1640 + 10% FBS Heat inactivated. After 2 hours wash with PBS to remove non-adherent cells and add your complete media ( RPMI 1640 + 10% FBS Heat inactivated). Then leave it for 12 hours (usually O/N) then wash with PBS and treat with your drug in Experimental media.
1. Isolate count plate (RPMI1640 ; 10% FBS-HI)
2. After 2h-3h: First wash with warm PBD (3 times),
3. O/N (RPMI1640 ; 10% FSB-HI),
4. Second wash with warm PBS (3 times)
5. Treat with drugs in the experimental media.
Just curious question.. How many cells you guys plate to get a better RNA in a 12 well plate?
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