人胞内细胞因子检测刺激方案
 |
| 人细胞因子 | 细胞来源 | 活化-极化培养 | 培养时间 | 再次刺激 | 胞内阻断剂 |
| G-CSF | PBMC | LPS (1 ug/mL) | 24hr | - | Monensin |
| GM-CSF | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Monensin |
| GIFN gamma | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Brefeldin A |
| IL-1 alpha | PBMC | LPS (1 ug/mL) | 24hr | - | Monensin |
| IL-1 beta | PBMC | LPS (100ng/mL) | 4hr | - | Brefeldin A |
| IL-1RA | PBMC | LPS (100ng/mL) | 24hr | - | Brefeldin A |
| IL-2 | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 4-6hr | - | Brefeldin A |
| IL-4 | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 4-6hr | - | Brefeldin A |
| IL-5 | CD4 | Th2 polarizing cultures | 6d | PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr) | Brefeldin A |
| IL-6 | PBMC | LPS (100ng/mL) | 24hr | - | Brefeldin A |
| IL-9 | CD4 | h2 polarizing culturesT | 6d | PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr) | Monensin |
| IL-10 | CD4 | Th2 polarizing cultures | 6d | PMA (50ng/ml) + Ionomycin (1ug/ml) (5hr) | Monensin |
| IL-12/ IL-23 (p40) | PBMC |
hIFN gamma (100ng/mL) (2hr)/LPS (100ng/mL) (22hr) | 2hr/22hr | - | Brefeldin A |
| IL-13 | CD4 | anti-CD3 (10ug/mL, immobilized) + anti-CD28 (2ug/mL, soluble) + IL-2 (10ng/mL) + IL-4 (20ng/mL) (2d); IL-2 (10ng/mL) + IL-4 (20ng/mL) (3d) | 2d/3d | PMA (5ng/ml) + Ionomycin (500ng/ml) (4hr) | Brefeldin A |
| IL-17A | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Monensin |
| IL-21 | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 4-7hr or 12-18hr | - | Brefeldin A |
| IL-22 | CD4 | Th17 polarizing cultures | 6d | PMA (50ng/ml) + Ionomycin (1ug/ml) (5hr) | Brefeldin A |
| IL-23 p19 | PBMC | hGM-CSF (40ng/mL) +hIL-4 (40ng/mL) | 6d | LPS (1ug/mL) (24hr) | Monensin |
| MCP-1/ CCL2 | PBMC | LPS (1ug/mL) | 24hr | - | Monensin |
| RANTES/ CCL5 | PBMC | LPS (1ug/mL) | 24hr | - | Monensin |
| TNF alpha | PBMC | PMA (30-50ng/mL)/Iono (1ug/mL) | 5hr | - | Brefeldin A |
| TNF beta | PBMC | Th1 polarizing cultures | 6d | PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr) | Monensin |
|
注： PBMC=外周血单核细胞peripheral blood mononuclear cell；PMA=佛波酯Phorbol Myristate Acetate; calcium ionophore; 钙离子载体 iono=离子霉素ionomycin; LPS=脂多糖Lipopolysaccharide; 4hr=4 hours culture; 2d=2 days culture. GolgiStop/GolgiPlug: 一种胞内阻断剂，可以用BFA/Monensin替代。
| |||||
 |
阅读原文：http://www.liankebio.com/TechServiceShow/articleID/2014060019.html
