Splenocyte Preperation
Harvest mouse spleen and prepare a single cell suspension.
Use slides or a syrings to push the spleen through a cell strainer.
Pellet the cells by centrifugation (350 x g); aspirate the supernatant.
Dilute the 10X Red Blood Cell Lysis Buffer to 1X working concentration with deionized water and resuspend the pellet in 5 ml of 1X Lysis Buffer.
Incubate on ice for 4-5 minutes with occasional shaking.
Stop the reaction by diluting the Lysis Buffer with 20-30 ml of 1X PBS.
Spin the cells (350 x g) and discard the supernatant.
Resuspend the pellet in the appropriate buffer Count cells, adjust density, and proceed with cell staining procedures.
