小鼠胞內細胞因子檢測刺激方案

瀏覽次數：7348　發布日期：2014-6-23　來源：本站　僅供參考，謝絕轉載，否則責任自負

小鼠細胞因子	細胞來源	活化-極化培養	培養時間	再次刺激	胞內阻斷劑
GM-CSF	mouse spleen	ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)	2d/3d	anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr	Brefeldin A
IFN-gamma	mouse spleen	ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)	2d/3d	anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr	Brefeldin A
IL-1 alpha	mouse PEC	mIFNγ (100ng/mL) (2hr)/LPS (100ng/mL)(22hr)	2hr/22hr	-	Brefeldin A
IL-1 beta	mouse PEC	LPS (100 ng/mL) (22hr)	22hr	-	Monensin
IL-2	mouse spleen	ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)	2d/3d	anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr	Brefeldin A
IL-4	mouse spleen	Th2 polarized	6d	PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr	Brefeldin A
IL-5	mouse splenic CD4	ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)	2d/3d	immobilized) + anti-CD28 (2ug/mL soluble) 5hr	Brefeldin A
IL-6	mouse PEC	LPS (100 ng/mL) (22hr)	22hr	-	Monensin
IL-10	mouse spleen	ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)	2d/3d	anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr	Brefeldin A
IL-12/IL-23 (p40)	mouse PEC	LPS (100 ng/mL) (22hr)	22hr	-	Brefeldin A
IL-13	mouse spleen	Th2 polarized	6d	PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr	Brefeldin A
IL-17A	mouse spleen	Th17 polarized	6d	PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr	Monensin
IL-17F	mouse spleen	Th17 polarized	6d	PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr	Monensin
IL-21	mouse spleen	Th17 polarized	9d	PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr	Monensin
IL-22	mouse spleen	Th17 polarized	12d	PMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr	Brefeldin A
IL-23 p19	mouse bone marrow	mGM-CSF (40 ng/mL)	8d	LPS (1 ug/mL) (24 hr)	Monensin
MCP-1/ CCL2	mouse PEC	LPS (100 ng/mL) (24hr)	24hr	-	Brefeldin A
TNF alpha	mouse spleen	ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)	2d/3d	anti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hr	Brefeldin A
說明：mouse PEC=mouse thioglycolate-elicited peritoneal macrophages腹腔巨噬細胞; ConA=刀豆素Concanavalin A; Iono=離子霉素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 結構的寡聚脫氧核苷酸。
來源：http://us.ebioscience.com/resources/best-protocols/flow-cytometry/flow-cytometry-intracellular-staining-quick-guide.htm

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