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 Proliferaton
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