Quick protocol guidelines for human cytokine bioassays are immediately below. Alternatively, see the quick protocol guidelines for mouse cytokine assays.
Human Cytokine Bioassay Quick Guide
| Cytokine | Indicator Cells | Cell Density (x105) | Incubation Time (hours) | Cytokine Top Conc. (ng/mL) | ED50 | Read-Out/Comments |
|---|---|---|---|---|---|---|
| Activin A | MPC-11 | 2 | 72 | 50 | ≤0.75-3 ng/mL | Inhibition of proliferation |
| B18R | L929 | 3.5 | 40 | 100 | ND50 ≤800pg/mL | neutralization of EMCV protection by rmIFNa2 |
| BMP-2 | C3H/10T1/2 | 3 | 1 week | 5000 | ≤0.25-1ug/mL | alkaline phosphatase induction |
| CCL19 (MIP-3b) | T-cells | 50 | 4 | 600 | max chemotaxis observed at ≤100-200ng/mL | Chemotaxis |
| CCL21 | PBMCs | 10 | 3 | 1200 | max chemotaxis observed at ≤ 200-400ng/mL | Chemotaxis |
| CCL4 (MIP-1b) | Monocytes | 2 | 4 | 225 | max chemotaxis observed at ≤25-75ng/mL | Chemotaxis |
| CCL7 (MCP-3) | PBMCs | 10 | 4 | 300 | max chemotaxis observed at ≤100-150ng/mL | Chemotaxis |
| CCL8 (MCP-2) | PBMCs | 10 | 4 | 450 | max chemotaxis observed at ≤100-150ng/mL | Chemotaxis |
| CD257 (BAFF, Blys) | Mouse splenocytes | 50 | 72 | 2000 | ≤125pg/mL | Mouse splenocyte survival assay |
| CXCL10 (IP-10) | T cells | 50 | 3 | 300 | max chemotaxis observed at ≤100-200ng/mL | Chemotaxis |
| EGF | HeLa | 0.5 | 72 | 20 | ≤1ng/mL | Proliferation |
| FGFb | Balb/c 3T3 | 1 | 72 | 50 | ≤0.1-1.25 ng/mL | Proliferation |
| Flt3-L | M1 | 3 | 72 | 250 | ≤5-15 ng/mL | induction of mIL-6 production, costim with 1ng/mL rmLIF |
| G-CSF | M-NFS-60 | 2 | 48 | 10 | ≤10-45 pg/mL | Proliferation |
| GM-CSF | TF-1 (ATCC CRL-2003) | 100 | 48 | 10 | ≤100pg/mL | Proliferation |
| HGF | 4MBr-5 (ATCC CCL-208) | 2 | 96 | 1000 | ≤50ng/mL | Proliferation |
| IFN-γ | A549 (ATCC CCL-185) | 3.5 | 40 | 10 | ≤40 pg/mL | EMCV protection |
| IL-1α | D10.G4.1 (ATCC TIB-224) | 5000 | 48 | 1 | ≤20-30pg/mL | Proliferation |
| IL-1β | D10.G4.1 (ATCC TIB-224) | 100 | 48 | 1 | ≤125pg/mL | Proliferation |
| IL-2 | CTLL-2 (ATCC TIB-214) | 2 | 24 | 10 | ≤175pg/mL | Proliferation |
| IL-3 | TF-1 (ATCC CRL-2003) | 2 | 48 | 10 | ≤0.6-0.15ng/mL | Proliferation |
| IL-4 | TF-1 (ATCC CRL-2003) | 2 | 48 | 10 | ≤175 pg/mL | Proliferation |
| IL-6 | TF-1 (ATCC CRL-2003) | 2 | 48 | 10 | ≤0.1ng/mL | Proliferation |
| IL-7 | Human PHA-treated PBMCs (3day) | 5 | 48 | 100 | ≤2000pg/mL | Proliferation |
| IL-8 | Human Neutrophils | 100 | 2 | 1000 | max chemotaxis observed at ≤10-100ng/mL | Chemotaxis |
| IL-10 | MC/9 (ATCC CRL-8306) | 2 | 48 | 10 | ≤1000 pg/mL | Proliferation |
| IL-12 p70 | Human PHA-treated PBMCs (3day) | 5 | 48 | 10 | ≤40-80pg/mL | Proliferation |
| IL-13 | TF-1 (ATCC CRL-2003) | 2 | 48 | 10 | ≤1000-2500pg/mL | Proliferation |
| IL-15 | CTLL-2 (ATCC TIB-214) | 3 | 48 | 10 | ≤100pg/mL | Proliferation |
| IL-17A | NHDF | 3 | 48 | 100 | ≤0.5-5ng/mL | Human IL-6 production |
| IL-17A/F | NIH/3T3 | 5 | 24 | 10,000 | ≤1500000pg/mL | Mouse IL-6 production |
| IL-17F | NIH/3T3 | 5 | 24 | 10,000 | ≤1500000pg/mL | Mouse IL-6 production |
| IL-21 | B9 | 3.5 | 48 | 10 | ≤25ng/mL | Proliferation |
| IL-22 | Colo205 | 10 | 24 | 1 | ≤50-100pg/mL | Human IL-10 Production |
| IL-23 | Mouse splenocytes | 50 | 24 | 100 | ≤2-3.5ng/mL | Mouse IL-17A Production |
| IL-27 | Mouse splenocytes | 10 | 72 | 1000 | ≤62.5-100 ng/mL | IL-27 mediated inhibition of mIL-2 production by mspl activated with CD3/CD28 |
| IL-28A | HepG2 | 3.5 | 40 | 100 | ≤1-5ng/mL | EMCV Protection |
| IL-29 | HepG2 | 3.5 | 40 | 10 | ≤0.5ng/mL | EMCV Protection |
| IL-33 | D10.G4.1 | 2 | 72 | 10 | ≤1000pg/mL | Proliferation |
| LIF | M1 | 3 | 48 | 100 | ≤15ng/mL | Mouse IL-6 Production |
| M-CSF | M-NFS-60 | 2 | 48 | 100 | ≤3ng/mL | Proliferation |
| PDGF-BB | Balb/c 3T3 | 1 | 72 | 500 | ≤30ng/mL | Proliferation |
| TGF-β1 | HT-2 (CRL-1841) | 2 | 48 | 10 | ≤40pg/mL | inhibition of mIL-4-induced proliferation |
| TGF-β2 | CTLL-2 | 2 | 72 | 50 | ≤0.3-1.5ng/mL | inhibition of mIL-4-induced proliferation |
| TGF-β3 | CTLL-2 | 2 | 48 | 5 | ≤0.03-0.125ng/mL | inhibition of mIL-4-induced proliferation |
| TNF-α | L929 | 3.5 | 24 | 10 | ≤80pg/mL | Cytotoxicity Assay |
| VEGF 121 | HUVEC | 2 | 72 | 1000 | ≤8-10ng/mL | Proliferation |
Mouse Cytokine Bioassay Quick Guide
| Cytokine | Indicator Cells | Cell Density (x105) | Incubation Time (hours) | Cytokine Top Conc. (ng/mL) | ED50 | Read-Out/Comments |
|---|---|---|---|---|---|---|
| CCL3 (MIP-1a) | Monocytes | 10 | 4 | 150 | max chemotaxis observed at ≤2-10ng/mL | Chemotaxis |
| CCL5 (RANTES) | Monocytes | 10 | 4 | 300 | max chemotaxis observed at ≤100ng/mL | Chemotaxis |
| Flt3 Ligand | M1 | 3 | 72 | 2000 | ≤50-250 ng/mL | Mouse IL-6 production, 1 ng/mL rmLIF co-stim |
| GM-CSF | MC/9 (ATCC CRL-8306) | 2 | 48 | 10 | ≤650pg/mL | Proliferation |
| HGF | mIMCD-3 | 2 | 48 | 100 | ≤6000-25000 pg/mL | Proliferation |
| HMGB1 | RAW264.7 | 3.5 | 24 | 105 | 3-12ug/mL | TNFa production, with 10 ug/mL of Polymycin B |
| IFNα2 | L929 | 3.5 | 40 | 10 | ≤0.1ng/mL | EMCV cell protection assay |
| IFNα4 | L929 | 3.5 | 40 | 1 | ≤75pg/mL | EMCV cell protection assay |
| IFN-γ | L929 | 3.5 | 40 | 10 | ≤175pg/mL | EMCV cell protection assay |
| IL-1α | D10.G4.1 (ATCC TIB-224) | 2 | 48 | 1 | ≤10 pg/mL | D10.G4.1 Proliferation |
| IL-1β | D10.G4.1 (ATCC TIB-224) | 3 | 48 | 1 | ≤10 pg/mL | D10.G4.1 Proliferation |
| IL-2 | CTLL-2 | 2 | 72 | 10 | ≤175pg/mL | Proliferation |
| IL-3 | MC/9 (ATCC CRL-8306) | 2 | 48 | 10 | ≤0.1ng/mL | Proliferation |
| IL-4 | CTLL-2 | 2 | 72 | 10 | ≤350pg/mL | Proliferation |
| IL-5 | TF-1 (ATCC TIB-214) | 2 | 48 | 10 | ≤150-625 pg/mL | Proliferation; Starve cells for 48-72 hrs prior |
| IL-6 | B9 | 2 | 48 | 0.1 | ≤5pg/mL | Proliferation; Starve cells for 48-72 hrs prior |
| IL-7 | hPBMC, 3 day PHA stimulated | 5 | 48 | 100 | ≤1600 pg/mL | Proliferation; PHA (5 ug/mL) as costimulator |
| IL-9 | TS-1 | 2 | 72 | 50 | ≤250-2000 pg/mL | Proliferation |
| IL-10 | MC/9 (ATCC CRL-8306) | 2 | 48 | 10 | ≤300 pg/mL | Proliferation; mIL-4 (5 pg/mL) as costimulator |
| IL-12 p70 | Splenocytes | 50 | 24 | 10 | ≤175pg/mL | Mouse IFN-γ production |
| IL-13 | TF-1 | 2 | 48 | 100 | ≤3.8ng/mL | Proliferation, Starve cells for 48-72 hrs prior |
| IL-15 | CTLL-2 (ATCC TIB-214) | 2 | 48 | 104 | ≤100ng/mL | Proliferation |
| IL-15R/IL-15 | CTLL-2 (ATCC TIB-214) | 2 | 48 | 250 | ≤4ng/mL | Proliferation |
| IL-17A | NIH/3T3 | 5 | 24 | 100 | ≤3ng/mL | Mouse IL-6 production |
| IL-17A/F | NIH/3T3 | 5 | 24 | 104 | ≤100ng/mL | Mouse IL-6 production |
| IL-17F | NIH/3T3 | 5 | 24 | 104 | ≤1250-2500 ng/mL | Mouse IL-6 production |
| IL-21 | B9 | 2 | 48 | 10 | ≤100-300 pg/mL | Proliferation; Starve cells for 48-72 hrs prior |
| IL-22 | Colo205 | 10 | 24 | 1 | ≤50-100 pg/mL | Human IL-10 production |
| IL-23 | Splenocytes | 50 | 24 | 10 | ≤625pg/mL | Mouse IL-17 production |
| IL-27 | Splenocytes | 10 | 72 | 100 | ≤3000 pg/mL | Inhibition of Mouse IL-2 secretion; immobilized @CD3, soluble @CD28 as costimulators |
| IL-33 | D10.G4.1 (ATCC TIB-224) | 2 | 72 | 1 | ≤30-100pg/mL | Proliferation |
| LIF | M1 | 3 | 48 | 50 | ≤200-750pg/mL | Mouse IL-6 production |
| M-CSF | M-NFS-60 | 2 | 48 | 250 | ≤10ng/mL | Proliferation |
| Noggin | C3H/10T1/2 | 3.5 | 1 week | 5000 | ≤150-300 ng/mL | inhibition of alkaline phosphatase by BMP-2 |
| SCF | MC/9 | 2 | 72 | 500 | ≤15000pg/mL | Proliferation |
| Sonic Hedgehog (SHH) | C3H/10T1/2 | 3 | 1 week | 2.5x104 | ≤0.5-3ug/mL | Alkaline phosphatase induction |
| TNF-α | L929 | 3.5 | 24 | 1 | ≤25pg/mL | L929 cytotoxicity assay |
| TSLP | Nag8/7 | 2 | 48 | 25 | ≤0.2ng/mL | Proliferation |
| VEGF 120 | HUVEC | 2 | 72 | 1000 | ≤8ng/mL | Proliferation |
仅供科研使用，不可用于诊断目的。