See below for intracellular staining quick guides for mouse and human cytokines as well as for non-cytokine proteins.

Mouse cytokines: intracellular staining quick guide for flow cytometry

Mouse cytokineCell sourceActivationIncubation timeRestimulationIntracellular blockAntibody
GM-CSFmouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AMP1-22E9
IFN-gammamouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AXMG1.2
IL-1 alphamouse PECmIFNγ (100ng/mL) (2hr)/LPS (100ng/mL)(22hr)2hr/22hr-Brefeldin AALF-161
IL-1 betamouse PEC LPS (100 ng/mL) (22hr)
IL-2mouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AJES6-5H4
IL-4mouse spleenTh2 polarized
6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hr
Brefeldin ABVD6-24G2,
IL-5mouse splenic CD4ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3dimmobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin ATRFK5
IL-6mouse PECLPS (100 ng/mL) (22hr)22hr-MonensinMP5-20F3
IL-10mouse spleenConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AJES5-16E3,
IL-12/IL-23 (p40)mouse PECLPS (100 ng/mL) (22hr)22hr-Brefeldin AC17.8
IL-13mouse spleenTh2 polarized6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrBrefeldin AeBio13A
IL-17Amouse spleenTh17 polarized
6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrMonensineBio17B7
IL-17Fmouse spleen
Th17 polarized
6dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrMonensineBio18F10 
IL-21mouse spleenTh17 polarized9dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrMonensinFFA21
IL-22mouse spleenTh17 polarized12dPMA (50 ng/mL) + Ionomycin (1 ug/mL) 5hrBrefeldin AIL22JOP
IL-23 p19mouse bone marrowmGM-CSF (40 ng/mL)8dLPS (1 ug/mL) (24 hr)Monensinfc23cpg
MCP-1/ CCL2mouse PECLPS (100 ng/mL) (24hr)
24hr-Brefeldin A2H5
TNF alphamouse spleen
ConA (3ug/mL) (2d)/IL-2 (20ng/mL)+IL-4 (20ng/mL) (3d)2d/3danti-CD3 (10ug/mL immobilized) + anti-CD28 (2ug/mL soluble) 5hrBrefeldin AMP6-XT22,

Annotations: mouse PEC=mouse thioglycolate-elicited peritoneal macrophages; ConA=Concanavalin A; Iono=Ionomycin; LPS=Lipopolysaccharide; PMA=Phorbol Myristate Acetate; 2d=2 day culture; 5hr=5 hour culture

Human cytokines: intracellular staining quick guide

Human cytokineCell sourceActivationIncubation timeRestimulationIntracellular blockAntibody
G-CSFPBMCLPS (1 ug/mL)24hr-Monensin8F5CSF
GM-CSFPBMCPMA (30-50ng/mL)/Iono (1ug/mL)5hr-MonensinBVD2-21C11
IFN gammaPBMCPMA (30-50ng/mL)/Iono (1ug/mL)5hr-Brefeldin A4S.B3
IL-1 alphaPBMCLPS (1 ug/mL)24hr-Monensin364/3B3-14, CRM8
IL-1 betaPBMCLPS (100ng/mL)4hr-Brefeldin ACRM56
IL-1RAPBMCLPS (100ng/mL)24hr-Brefeldin ACRM17
IL-2PBMCPMA (30-50ng/mL)/Iono (1ug/mL)4-6hr-Brefeldin AMQ1-17H12
IL-4PBMCPMA (30-50ng/mL)/Iono (1ug/mL)4-6hr-Brefeldin A8D4-8
IL-5CD4Th2 polarizing cultures
PMA (50ng/mL) + Ionomycin (1ug/mL) (5hr)Brefeldin ATRFK5, JES1-5A10
IL-6PBMCLPS (100ng/mL)24hr-Brefeldin AMQ2-13A5
IL-9CD4Th2 polarizing cultures6dPMA (50ng/mL) + Ionomycin (1ug/mL) (5hr)MonensinMH9A4
IL-10CD4Th2 polarizing cultures6dPMA (50ng/ml) + Ionomycin (1ug/ml) (5hr)
IL-12/ IL-23 (p40)PBMChIFN gamma (100ng/mL) (2hr)/LPS (100ng/mL) (22hr)2hr/22hr-Brefeldin AC8.6
IL-13CD4anti-CD3 (10ug/mL, immobilized) + anti-CD28 (2ug/mL, soluble) + IL-2 (10ng/mL) + IL-4 (20ng/mL) (2d); IL-2 (10ng/mL) + IL-4 (20ng/mL) (3d)2d/3dPMA (5ng/ml) + Ionomycin (500ng/ml) (4hr)Brefeldin APVM13-1
IL-17APBMCTh17 polarizing cultures
6dPMA (50ng/ml) + Ionomycin (1ug/ml) (5hr)Brefeldin AeBio64CAP17,


IL-21PBMCPMA (30-50ng/mL)/Iono (1ug/mL)4-7hr or 12-18hr-Brefeldin AeBio3A3-N2
IL-22CD4Th17 polarizing cultures6dPMA (50ng/ml) + Ionomycin (1ug/ml) (5hr)Brefeldin AIL22JOP
IL-23 p19PBMChGM-CSF (40ng/mL) +hIL-4 (40ng/mL)6dLPS (1ug/mL) (24hr)Monensin23dcdp


RANTES/ CCL5PBMCLPS (1ug/mL)24hr-MonensinVL1
TNF alphaPBMCPMA (30-50ng/mL)/Iono (1ug/mL)5hr-Brefeldin AMAb11
TNF betaPBMCTh1 polarizing cultures
6dPMA (50ng/mL) + Ionomycin (1ug/mL) (5hr)Monensin359-81-11

Annotations: Iono=Ionomycin; PMA=Phorbol Myristate Acetate; LPS=Lipopolysaccharide; 2d=2 day culture; 5hr=5 hour culture; LPS for activation of human PBMC obtained from Sigma (#L-8274)

Non-cytokine proteins: intracellular staining quick guide

Mouse/Rat Bcl-210C4
Mouse CTLA-4 (CD152)UC10-4B9
Human CTLA-4 (CD152)14D3
Human/Mouse/Rat Cytochrome C6H2
Human Foxp3PCH101
Mouse Foxp3FJK-16s
Mouse/Human Granzyme BeBioGrB
House Langerin (CD207)eBioRMUL.2
Human NanoghNanog.1
Human PCNA (Proliferating Cell Nuclear Antigen)PC10
Mouse PerforineBioOMAK-D
Human PerforindG9
ROR gamma 
Mouse SLP-76MS76
Human SLP-76HS76
Human/Mouse T-beteBio4B10 (4B10, 4-B10)
Human TLR3TLR3.7
Mouse TLR9M9.D6
Human TLR9eB72-1665
Mouse/Human ZAP-701E7.2

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