Cytokine Bioassays Quick Guide | Thermo Fisher Scientific

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

For Research Use Only. Not for use in diagnostic procedures.

Cytokine Bioassays Quick Guides

Human Cytokine Bioassay Quick Guide

Mouse Cytokine Bioassay Quick Guide