| Cytokine/Chemokine | Clone(s) | Target Cells | Stimulation | Stimulation period | Protein transport blocker | Surface marker |
|---|---|---|---|---|---|---|
| IL-1α | 364-3B3-14 | PBMCs | LPS (100 ng/ml) | 4-6 hours | monensin or brefeldin A | CD14* |
| IL-1β | JK1B-1 | PBMCs | LPS (100 ng/ml) | 4-6 hours | monensin or brefeldin A | CD14* |
| IL-2 | MQ1-17H12 | PBMCs | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD3 |
| IL-3 | BVD3-1F9 | Purified CD4+ T cells stimulated with immobilized anti-CD3 (10 μg/ml) + soluble anti-CD28 (5 μg/ml) + IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 2 days, wash and then culture in IL-2 + IL-4 for 3-4 days | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-4 | MP4-25D2 or 8D4-8 | PBMCs | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-5 | JES1-39D10 | Purified CD4+ T cells stimulated with immobilized anti-CD3 (10 μg/ml) + soluble anti-CD28 (5 μg/ml) + IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 2 days, wash and then culture in IL-2 + IL-4 for 3-4 days | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-6 | MQ2-13A5 | PBMCs | LPS (100 ng/ml) | 4-6 hours | monensin or brefeldin A | CD14* |
| IL-8 | JK8-1 | PBMCs | LPS (100 ng/ml) | 4-6 hours | monensin or brefeldin A | CD14* |
| IL-9 | MH9A4 | Purified CD4+ T cells stimulated with immobilized anti-CD3 (10 μg/ml) + soluble anti-CD28 (5 μg/ml) + IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 2 days, wash and then culture in IL-2 + IL-4 for 3-4 days | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-10 | JES3-9D7 or JES3-19F1 | PBMCs | LPS (100 ng/ml) | 18-24 hours | monensin or brefeldin A | CD14* |
| IL-12 p40 | C11.5 | PBMCs | IFN-γ (100 ng/ml) prime for 2 hours, then add LPS (1 μg/ml) | 18-24 hours | monensin or brefeldin A | CD14* |
| IL-13 | JES10-5A2 | Purified CD4+ T cells stimulated with immobilized anti-CD3 (10 μg/ml) + soluble anti-CD28 (5 μg/ml) + IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 2 days, wash and then culture in IL-2 + IL-4 for 3-4 days | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-17A | BL168 | PBMCs | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-21 | 3A3-N2 or 4BG1 | PBMCS | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| IL-22 | BG/IL22 | PBMCs | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD4* |
| GM-CSF | BVD2-21C11 | PBMCs stimulated with soluble anti-CD3 (UCHT1, 1 μg/ml) + soluble anti-CD28 (5 μg/ml) for 2 days, wash and then culture in IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 3-4 days | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD3 |
| IFN-γ | 4S.B3 or B27 | PBMCs | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD3 |
| MCP-1 | 5D3-F7 or 2H5 | PBMCs | LPS (100 ng/ml) | 18-24 hours | monensin or brefeldin A | CD14* |
| TNF-α | MAB11 | PBMCs | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD3 |
| TNF-β | 359-81-11 | PBMCs stimulated with soluble anti-CD3 (UCHT1, 1 μg/ml) + soluble anti-CD28 (5 μg/ml) for 2 days, wash and then culture in IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 3-4 days | PMA (50 ng/ml) plus ionomycin (1 μg/ml) | 4-6 hours | monensin or brefeldin A | CD3 |