Human IL-27 ELISA Kit PicoKine®

Interleukin-27 (IL-27) is a heterodimeric cytokine belonging to the IL-12 family that is composed of two subunits; Epstein-Barr virus (EBV) -induced gene 3 (EBI3) (also known as IL-27B) and IL27-p28 (known as IL-30) . IL-27 is produced by antigen-presenting cells.IL-27 plays an important function in regulating the activity of B- and T-lymphocytes. IL-27, a potent inhibitor of T (H) -17 cell development, may be a useful target for treating inflammatory diseases mediated by these cells.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 266 272 277 241 264 13.83 5.2% Sample 2 1556 1617 1541 1521 1558 35.84 2.3% Sample 3 5164 5696 5356 5370 5396 191.11 3.5% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 266 272 277 241 264 13.83 5.2% Sample 1 266 272 277 241 264 13.83 5.2% Sample 2 1556 1617 1541 1521 1558 35.84 2.3% Sample 2 1556 1617 1541 1521 1558 35.84 2.3% Sample 3 5164 5696 5356 5370 5396 191.11 3.5% Sample 3 5164 5696 5356 5370 5396 191.11 3.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 266 1556 5164 259 1491 5174 Standard deviation 14.63 716.05 256.2% 16.05 90.95 284.57 CV (%) 5.5% 5% 5% 6.2% 6.1% 5.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 266 1556 5164 259 1491 5174 Standard deviation 14.63 716.05 256.2% 16.05 90.95 284.57 CV (%) 5.5% 5% 5% 6.2% 6.1% 5.5% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 266 1556 5164 259 1491 5174 Mean (pg/ml) 266 1556 5164 259 1491 5174 Standard deviation 14.63 716.05 256.2% 16.05 90.95 284.57 Standard deviation 14.63 716.05 256.2% 16.05 90.95 284.57 CV (%) 5.5% 5% 5% 6.2% 6.1% 5.5% CV (%) 5.5% 5% 5% 6.2% 6.1% 5.5%

Associates with EBI3 to form the IL-27 interleukin, a heterodimeric cytokine which functions in innate immunity. IL-27 has pro- and anti-inflammatory properties, that can regulate T- helper cell development, suppress T-cell proliferation, stimulate cytotoxic T-cell activity, induce isotype switching in B-cells, and that has diverse effects on innate immune cells. Among its target cells are CD4 T-helper cells which can differentiate in type 1 effector cells (TH1), type 2 effector cells (TH2) and IL17 producing helper T-cells (TH17) . It drives rapid clonal expansion of naive but not memory CD4 T-cells. It also strongly synergizes with IL-12 to trigger interferon-gamma/IFN-gamma production of naive CD4 T-cells, binds to the cytokine receptor WSX-1/TCCR which appears to be required but not sufficient for IL-27-mediated signal transduction. IL-27 potentiate the early phase of TH1 response and suppress TH2 and TH17 differentiation. It induces the differentiation of TH1 cells via two distinct pathways, p38 MAPK/TBX21- and ICAM1/ITGAL/ERK-dependent pathways. It also induces STAT1, STAT3, STAT4 and STAT5 phosphorylation and activates TBX21/T-Bet via STAT1 with resulting IL12RB2 up- regulation, an event crucial to TH1 cell commitment. It suppresses the expression of GATA3, the inhibitor TH1 cells development. In CD8 T-cells, it activates STATs as well as GZMB. IL-27 reveals to be a potent inhibitor of TH17 cell development and of IL-17 production. Indeed IL27 alone is also able to inhibit the production of IL17 by CD4 and CD8 T-cells. While IL-27 suppressed the development of proinflammatory Th17 cells via STAT1, it inhibits the development of anti-inflammatory inducible regulatory T-cells, iTreg, independently of STAT1. IL-27 has also an effect on cytokine production, it suppresses proinflammatory cytokine production such as IL2, IL4, IL5 and IL6 and activates suppressors of cytokine signaling such as SOCS1 and SOCS3. Apart from suppression of cytokine production, IL-27 also antagonizes the effects of some cytokines such as IL6 through effects on T- cells. Another important role of IL-27 is its antitumor activity as well as its antiangiogenic activity with activation of production of antiangiogenic chemokines such as IP-10/CXCL10 and MIG/CXCL9. In vein endothelial cells, it induces IRF1/interferon regulatory factor 1 and increase the expression of MHC class II transactivator/CIITA with resulting up-regulation of major histocompatibility complex class II. IL-27 also demonstrates antiviral activity with inhibitory properties on HIV-1 replication. .

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.062 0.146 0.232 0.383 0.651 1.083 1.685 2.178 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.062 0.146 0.232 0.383 0.651 1.083 1.685 2.178 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.062 0.146 0.232 0.383 0.651 1.083 1.685 2.178 O.D. 0.062 0.146 0.232 0.383 0.651 1.083 1.685 2.178

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.