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Version: 3 Revision Date: 09/03/2014 Recombinant Human IL-35-Fc fusion protein (ELISA Std.) Catalog# / Size 578502 / 10 µg Other Names IL-35 subunits: IL-12 subunit alpha (p35), IL-27 subunit beta (EBI3) Ave. Rating ★★★★★ 0 reviews Description IL-35 is a newly described cytokine comprised of the IL-12α chain (p35) and the Epstein-Barr virus-induced gene product 3 (EBI-3). IL-35 belongs to the IL-12 family which includes IL-12, IL-23, IL-27, and IL-35. These cytokines are heterodimeric proteins comprised of an α chain (p19, p28, or p35) and a β chain (p40 or Ebi3). Mouse IL-35-Fc fusion protein enhances the proliferation of mouse Treg cells, while inhibiting the development of Th17 cells. Human IL-35 can suppress the proliferation of umbilical cord-derived human CD4 T conventional cells, to a similar extent as activated Tregs. In addition, activation of human CD4 T conventional cells with anti-CD3 and anti-CD28 in the presence of IL-35 induces the expression of IL-35 in these cells. Interestingly, activated T conventional cells treated with IL-35 acquire a regulatory phenotype (iTreg35) that suppress via IL-35. Most recent data suggest that IL-35 secreted by ICOS+ Tregs can suppress IL-17 production and reverse established IL-17-dependent allergic airways disease in mice. Reactivity Human Source Human IL-35 consists of two subunits linked via a 3x (SGGGG) bridge: EBI3 (Accession# NM_005755.2) and p35 (Accession# NM_000882.3). Human IL-35 was expressed in 293E cells as a human Fc-fusion protein (SP-EBI3-linker-p35-hFc). Molecular Mass The 762 amino acid recombinant protein has a predicted molecular weight of 84.4 kD. The non -reduced migrates at approximately 200 kD, and the DTT-reduced protein migrates at approximately 100 kD by SDS-PAGE. Purity >98%, as determined by Coomassie stained SDS-PAGE. Formulation 0.22 µm filtered protein solution is in PBS. Endotoxin Level Less than 0.01 ng per µg cytokine as determined by the LAL method. Preparation 10 µg size is bottled at 200 µg/mL. Storage & Handling Unopened vial can be stored between 2°C and 8°C for one month, at -20°C for six months, or at -70°C for one year. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored from -20°C to -70°C. Stock solutions can also be prepared at 50-100 µg/mL in sterile buffer (PBS, HPBS, DPBS, or EBSS) containing carrier protein such as 0.2-1% BSA or HSA and stored in working aliquots at -20°C to -70°C. Avoid repeated freeze/thaw cycles. Application ELISA Antigen Details Structure Heterodimer Distribution Regulatory T cells (Treg), CD4+ CD25+ Foxp3+ , CD4+ CD25+ , Foxp3- regulatory T cells. Function IL-35 suppresses T effector cell proliferation in vitro and induces a new subpopulation of Tregs (iTregs35). IL-35-Fc fusion protein enhances the proliferation of mouse Treg cells. Interaction between Tregs and T effector cells induces IL-35 production. Interaction T conventional cells, T regs. Ligand Receptor Unknown Antigen References 1. 2. 3. 4. 5. 6. Collison LW, et al. 2007. Nature 450:566. Niedbala W, et al. 2007. Eur. J. Immunol. 37:3021. Collison LW and Vignali DA. 2008. Immunol. Rev. 226:248. Ito T, el al. 2008. Immunity 28:870. Collison LW, et al. 2010. Nat. Immunol. 12:1093. Whitehead GS, et al. 2012. J. Allergy Clin. Immunol. 129:207. Gene ID 3592 10148 UniProt View information about IL-35 on UniProt.org Documentation Technical data sheet Certificate of Analysis Safety Data Sheet Related Protocols Sandwich ELISA Protocol Related FAQs How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg? Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL) Related Categories Cytokines & Chemokines > RecombinantProteins > Human > IL-35 > Cytokines & Chemokines > Human Cytokine & Chemokine Research > Recombinant Proteins > IL-35 > Antibodies and More > Cell Biology > Signaling/Signaling Intermediates > IL-35 > For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products. *These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use. BioLegend Inc., 9727 Pacific Heights Blvd, San Diego, CA 92121 www.biolegend.com Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587