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Isolation of murine splenocytes: Difference between revisions
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Isolation of murine splenocytes (view source)
Revision as of 21:03, 25 September 2006
, 25 September 2006→Protocol A
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#Spin cells at 800xg for 3 minutes. | #Spin cells at 800xg for 3 minutes. | ||
#Discard supernatant and resuspend pellet in 1mL ACK lysis buffer. Incubate at RT for 5-10 minutes. Add 9mL DMEM-10 and spin as before. | #Discard supernatant and resuspend pellet in 1mL ACK lysis buffer. Incubate at RT for 5-10 minutes. Add 9mL DMEM-10 and spin as before. | ||
#Discard supernatant and resuspend pellet in 3mL DMEM-10. | #Discard supernatant and resuspend pellet in 3mL DMEM-10, discarding any dead cell mass. | ||
#Count cells (dilute 10μL cell suspension in trypan blue, and count with hemcytometer). | #Count cells (dilute 10μL cell suspension in trypan blue, and count with hemcytometer). | ||
