小鼠胞内细胞因子检测刺激方案

联科生物

2014/06/23 11:20

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小鼠细胞因子细胞来源活化-极化培养培养时间再次刺激胞内阻断剂GM-CSFmouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AIFN-gammamouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AIL-1 alphamouse PECmIFNγ (100ng/mL) (2hr)/LPS (100ng/mL)(22hr)2hr/22hr-Brefeldin AIL-1 betamouse PECLPS (100 ng/mL) (22hr)22hr-MonensinIL-2mouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin A IL-4mouse spleenTh2 polarized6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrBrefeldin AIL-5mouse splenic CD4ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3dimmobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin A IL-6mouse PECLPS (100 ng/mL) (22hr)22hr-MonensinIL-10mouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AIL-12/IL-23 (p40)mouse PECLPS (100 ng/mL) (22hr)22hr-Brefeldin A IL-13mouse spleenTh2 polarized6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrBrefeldin AIL-17Amouse spleenTh17 polarized6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrMonensinIL-17F mouse spleenTh17 polarized6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrMonensinIL-21mouse spleenTh17 polarized9dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrMonensinIL-22mouse spleenTh17 polarized12dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrBrefeldin AIL-23 p19mouse bone marrowmGM-CSF (40 ng/mL)8dLPS (1 ug/mL) (24 hr)MonensinMCP-1/ CCL2mouse PECLPS (100 ng/mL) (24hr)24hr-Brefeldin ATNF alphamouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin A说明：mouse PEC=mouse thioglycolate-elicited peritoneal macrophages腹腔巨噬細胞; ConA=刀豆素Concanavalin A; Iono=离子霉素Ionomycin; LPS=脂多糖Lipopolysaccharide; PMA=佛波酯Phorbol Myristate Acetate; 2d=2 day culture; 5hr=5 hour culture；CpG ODN 2395=未甲基化的含有CpG 结构的寡聚脱氧核苷酸。来源：http://us.ebioscience.com/resources/best-protocols/flow-cytometry/flow-cytometry-intracellular-staining-quick-guide.htm