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

CytokineIndicator CellsCell Density (x105)Incubation Time (hours)Cytokine Top Conc. (ng/mL)ED50 Read-Out/Comments
Activin A MPC-1127250≤0.75-3 ng/mLInhibition of proliferation
B18RL929 3.540100ND50 ≤800pg/mLneutralization of EMCV protection by rmIFNa2
BMP-2C3H/10T1/231 week5000≤0.25-1ug/mLalkaline phosphatase induction
CCL19 (MIP-3b)T-cells504600max chemotaxis observed
at ≤100-200ng/mL
Chemotaxis
CCL21PBMCs1031200max chemotaxis observed
at ≤ 200-400ng/mL
Chemotaxis
CCL4 (MIP-1b)Monocytes24225max chemotaxis observed
at ≤25-75ng/mL
Chemotaxis
CCL7 (MCP-3)PBMCs104300max chemotaxis observed
at ≤100-150ng/mL
Chemotaxis
CCL8 (MCP-2)PBMCs104450max chemotaxis observed
at ≤100-150ng/mL
Chemotaxis
CD257 (BAFF, Blys)Mouse splenocytes 50722000≤125pg/mLMouse splenocyte survival assay
CXCL10 (IP-10)T cells503300max chemotaxis observed
at ≤100-200ng/mL
Chemotaxis
EGFHeLa0.57220≤1ng/mLProliferation
FGFbBalb/c 3T317250≤0.1-1.25 ng/mLProliferation
Flt3-LM1372250≤5-15 ng/mLinduction of mIL-6 production, costim with 1ng/mL rmLIF
G-CSFM-NFS-6024810≤10-45 pg/mLProliferation
GM-CSF TF-1 (ATCC CRL-2003) 1004810≤100pg/mLProliferation
HGF 4MBr-5 (ATCC CCL-208) 2961000≤50ng/mLProliferation
IFN-γ A549 (ATCC CCL-185) 3.54010≤40 pg/mLEMCV protection
IL-1α D10.G4.1 (ATCC TIB-224) 5000481≤20-30pg/mLProliferaton
IL-1β D10.G4.1 (ATCC TIB-224) 100481≤125pg/mLProliferation
IL-2 CTLL-2 (ATCC TIB-214) 22410≤175pg/mLProliferation
IL-3 TF-1 (ATCC CRL-2003) 24810≤0.6-0.15ng/mLProliferation
IL-4 TF-1 (ATCC CRL-2003) 24810≤175 pg/mLProliferation
IL-6 TF-1 (ATCC CRL-2003) 24810≤0.1ng/mLProliferation
IL-7 Human PHA-treated PBMCs (3day) 548100≤2000pg/mLProliferation
IL-8 Human Neutrophils 10021000max chemotaxis observed
at ≤10-100ng/mL
Chemotaxis
IL-10 MC/9 (ATCC CRL-8306) 24810≤1000 pg/mLProliferation
IL-12 p70 Human PHA-treated PBMCs (3day) 54810≤40-80pg/mLProliferation
IL-13 TF-1 (ATCC CRL-2003) 24810≤1000-2500pg/mLProliferation
IL-15 CTLL-2 (ATCC TIB-214) 34810≤100pg/mLProliferation
IL-17A NHDF 348100≤0.5-5ng/mLHuman IL-6 production 
IL-17A/FNIH/3T352410,000≤1500000pg/mLMouse IL-6 production 
IL-17FNIH/3T352410,000≤1500000pg/mLMouse IL-6 production 
IL-21 B93.54810≤25ng/mLProliferation
IL-22 Colo20510241≤50-100pg/mLHuman IL-10 Production
IL-23 Mouse splenocytes 5024100≤2-3.5ng/mLMouse IL-17A Production
IL-27 Mouse splenocytes 10721000≤62.5-100 ng/mLIL-27 mediated inhibition of
mIL-2 production by mspl activated with CD3/CD28
IL-28AHepG23.540100≤1-5ng/mLEMCV Protection
IL-29HepG23.54010≤0.5ng/mLEMCV Protection
IL-33 D10.G4.1 27210≤1000pg/mLProliferation
LIFM1348100≤15ng/mLMouse IL-6 Production
M-CSFM-NFS-60248100≤3ng/mLProliferation
PDGF-BBBalb/c 3T3172500≤30ng/mLProliferation
TGF-β1HT-2 (CRL-1841) 24810 ≤40pg/mLinhibition of mIL-4-induced
proliferation
TGF-β2CTLL-227250≤0.3-1.5ng/mLinhibition of mIL-4-induced
proliferation
TGF-β3CTLL-22485≤0.03-0.125ng/mLinhibition of mIL-4-induced
proliferation
TNF-α L9293.52410≤80pg/mLCytotoxicity Assay
VEGF 121HUVEC2721000 ≤8-10ng/mL Proliferation

Mouse Cytokine Bioassay Quick Guide

CytokineIndicator CellsCell Density (x105)Incubation Time (hours)Cytokine Top Conc. (ng/mL)ED50 Read-Out/Comments
CCL3 (MIP-1a)
Monocytes104150max chemotaxis observed at ≤2-10ng/mLChemotaxis
CCL5 (RANTES)Monocytes104300max chemotaxis observed at ≤100ng/mLChemotaxis
Flt3 LigandM13722000≤50-250 ng/mLMouse IL-6 production, 1 ng/mL rmLIF co-stim
GM-CSF
MC/9 (ATCC CRL-8306)24810≤650pg/mLProliferation
HGF
mIMCD-3248100≤6000-25000 pg/mLProliferation
HMGB1
RAW264.73.5241053-12ug/mLTNFa production, with 10 ug/mL of Polymycin B
IFNα2L9293.54010≤0.1ng/mLEMCV cell protection assay
IFNα4L9293.5401≤75pg/mLEMCV cell protection assay
IFN-γL9293.54010≤175pg/mLEMCV cell protection assay
IL-1αD10.G4.1 (ATCC TIB-224)2481≤10 pg/mLD10.G4.1 Proliferation
IL-1βD10.G4.1 (ATCC TIB-224)3481≤10 pg/mLD10.G4.1 Proliferation
IL-2CTLL-227210≤175pg/mLProliferation
IL-3MC/9 (ATCC CRL-8306)24810≤0.1ng/mLProliferation
IL-4CTLL-227210≤350pg/mLProliferation
IL-5
TF-1 (ATCC TIB-214)
24810≤150-625 pg/mLProliferation; Starve cells for 48-72 hrs prior
IL-6B9
2480.1≤5pg/mLProliferation; Starve cells for 48-72 hrs prior
IL-7hPBMC, 3 day PHA stimulated
548100≤1600 pg/mLProliferation; PHA (5 ug/mL) as costimulator
IL-9
TS-127250≤250-2000 pg/mLProliferation
IL-10
MC/9 (ATCC CRL-8306)24810≤300 pg/mLProliferation; mIL-4 (5 pg/mL) as costimulator
IL-12 p70
Splenocytes502410≤175pg/mLMouse IFN-γ production
IL-13
TF-1
248100≤3.8ng/mLProliferation, Starve cells for 48-72 hrs prior
IL-15
CTLL-2 (ATCC TIB-214)
248104≤100ng/mLProliferation
IL-15R/IL-15
CTLL-2 (ATCC TIB-214)
248250≤4ng/mLProliferation
IL-17A
NIH/3T3524100≤3ng/mLMouse IL-6 production
IL-17A/FNIH/3T3524104≤100ng/mLMouse IL-6 production
IL-17FNIH/3T3
524104≤1250-2500 ng/mLMouse IL-6 production
IL-21B924810≤100-300 pg/mLProliferation; Starve cells for 48-72 hrs prior
IL-22
Colo20510241≤50-100 pg/mLHuman IL-10 production
IL-23Splenocytes502410≤625pg/mLMouse IL-17 production
IL-27Splenocytes1072100≤3000 pg/mLInhibition of Mouse IL-2 secretion; immobilized @CD3, soluble @CD28 as costimulators
IL-33D10.G4.1 (ATCC TIB-224)2721≤30-100pg/mLProliferation
LIFM1
34850≤200-750pg/mLMouse IL-6 production
M-CSFM-NFS-60248250≤10ng/mLProliferation
NogginC3H/10T1/23.51 week5000≤150-300 ng/mLinhibition of alkaline phosphatase by BMP-2
SCFMC/9272500≤15000pg/mLProliferation
Sonic Hedgehog (SHH)C3H/10T1/231 week2.5x104≤0.5-3ug/mLAlkaline phosphatase induction
TNF-α
L9293.5241≤25pg/mLL929 cytotoxicity assay
TSLPNag8/724825≤0.2ng/mLProliferation
VEGF 120HUVEC2721000≤8ng/mLProliferation