小鼠胞内细胞因子检测刺激方案
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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-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 |
| 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 | |
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阅读原文：http://www.liankebio.com/TechServiceShow/articleID/2014060023.html
