Interleukins Focus: IL-1 Family & Other Newcomers

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1 International Edition THE EXERT FOR INTERLEUKINS Interleukins Focus: IL-1 Family & Other Newcomers The interleukin-1 (IL-1) family comprises 11 proteins that transmit their signals through 5 receptors: IL-1RI, IL-18R, IL-1Rrp2, ST2 and SIGGIR. They probably arose from an ancestral gene, since they possess similar structures. This family is categorized into subfamilies: the IL-1 subfamily consists of IL-1, IL-1 and IL-33. IL-18 and IL-37 belong to the IL-18 subfamily. IL-36, and (previously called IL-1F6, IL-1F8, IL-1F9, respectively) as well as IL-36Ra (IL-1F5) belong to the IL-36 subfamily. IL-38 (previously called IL-1F8) likely belongs to this IL-36 subfamily. Although all members are found in the extracellular space, only the IL-1 receptor antagonist (IL-1Ra) has a classical signal peptide for secretion through the endoplasmic reticulum/golgi pathway and therefore cannot be categorized into these subfamilies. All other members are first synthesized as a precursor without a defined signal peptide for processing and secretion. While most of these cytokines are biologically active as full-length molecules, activation and secretion of IL-1 and IL-18 requires inflammasome/ caspase-1-dependent processing. Other IL-1 family cytokines do not require caspase-1 cleavage for activation but may undergo some form of protease processing at their N-terminal ends, which leads to more potent proteins. IL-1 family members act on innate immune cells and play a central role in the activation of inflammation and in the first steps of innate immunity against pathogens or tissue damage, associated with acute and chronic inflammation. SELECTED REVIEWS: The IL-1 family: regulators of immunity: J.E. Sims & D.E. Smith; Nat. Rev. Immunol. 10, 89 (2010) New insights in the immunobiology of IL-1 family members: F.L. van de Veerdonk & M.G. Netea; Front. Immunol. 4, 167 (2013) NAME FAMILY NAME RECETOR CO-RECETOR ROERTY IL-1 IL-1F1 IL-1RI IL-1RAc roinflammatory IL-1 IL-1F2 IL-1RI IL-1RAc roinflammatory IL-1Ra IL-1F3 IL-1RI NA Antagonist for IL-1, IL-1 IL-18 IL-1F4 IL-18R IL-18R roinflammatory IL-36Ra IL-1F5 IL-1Rrp2 NA Antagonist for IL-36, IL-36, IL-36 IL-36 IL-1F6 IL-1Rrp2 IL-1RAc roinflammatory IL-37 IL-1F7 SIGGIR, IL-18R Unknown Anti-inflammatory IL-36 IL-1F8 IL-1Rrp2 IL-1RAc roinflammatory IL-36 IL-1F9 IL-1Rrp2 IL-1RAc roinflammatory IL-38 IL-1F10 Unknown Unknown Unknown IL-33 IL-1F11 ST2 IL-1RAc Th2 responses, roinflammatory Contents AGE IL-33 An Endogenous Danger Signal 2-3 IL-36,, roinflammatory Cytokines & IL-36Ra 4 IL-37 An Anti-inflammatory Cytokine 5 IL-38 A ro- or Anti-inflammatory Cytokine? 6 IL-6 otent Hybridoma Growth Factor 6 Other Interleukin Antibodies & roteins 7 IL-2 Superkine 7 IL-35 & The IL-12 Family Backcover

2 IL-33 [IL-1F11] An Endogenous Danger Signal IL-33 (also called DVS27 or nuclear factor from high endothelial venules (NF-HEV)), a member of the IL-1 family is a multifunctional proinflammatory cytokine. IL-33 was originally discovered as a nuclear protein expressed in high endothelial venules. Full-length IL-33 (pro-il-33) has a chromatin-binding domain at its N-terminus and is located in the nucleus where its function is not well characterized. ro-il-33 does not possess any signal peptide and may be secreted through unconventional secretory mechanisms or following cell damage and necrosis. Secreted pro-il-33 is cleaved by neutrophil serine proteases, cathepsin G and elastase that generate mature forms. Both pro-il-33 and mature forms of IL-33 can bind to the heteromeric receptor ST2/IL-1R accessory protein (IL-1RAc), but the mature forms are more active than the proform. Secreted IL-33 functions as an endogenous danger signal/alarmin and triggers inflammation in an autocrine or paracrine fashion. IL-33 upon receptor binding drives cytokine production in type 2 innate lymphoid cells (ILCs) (natural helper cells, nuocytes), T-helper (Th) 2 lymphocytes, mast cells, basophils, eosinophils, invariant natural killer T (inkt) cells and natural killer (NK) cells. IL-33 is involved in different diseases such as sepsis, asthma, anaphylaxia, rheumatoid arthritis, nervous system diseases, ulcerative colitis, allergy, infections, obesity and cardiovascular diseases. In healthy human serum almost no IL-33 can be detected, whereas serum IL-33 levels significantly increase in chronic active inflammation, like rheumatoid arthritis. SELECTED REVIEWS: Mechanisms of IL-33 processing and secretion: differences and similarities between IL-1 family members: E. Lefrancais & C. Cayrol; Eur. Cytokine Netw. 23, 120 (2012) Role of interleukin-33 in innate-type immune cells in allergy: S. Nakae, et al.; Allergol. Int. 62, 13 (2013) Translocation IL-33? Transcription NUCLEUS IL-1RAc MYD88 TRAF6 IKK DD IκB p50 p65 Ac IL-33 ST2 TIR Domain IRAK4 IRAK1/2 TRAF6 TAB1/2 TAK1 IL-33 roteasomal Degradation IL-1β, IL-3, IL-6, CXCL2, CCL2, CCL3 SIGGIR/TIR8 ERK p38 JNK A-1 Soluble ST2 CYTOLASM MAKKs IL-5, IL-13, CCL5, CCL17, CCL24 FIGURE: IL-33 Signaling athway THE STANDARD Specific Interleukin-33 ELISA Kits for Human or Mouse Interleukin-33 Detection Optimized for detection in cell culture supernatants, serum or plasma. Specific and sensitive for the measurement of natural and recombinant human or mouse interleukin-33. Reproducible results with low inter- and intra-assay variation. IL-33 (human) ELISA Kit AG-45A-0033EK-KI01 AG-45A-0033T-KI01 Twinlex 2 x AG-45A-0033-KI01 entalex 5 x SECIES REACTIVITY: Human 5 pg/ml 16 to 1000 pg/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Culture Supernatant, lasma, Serum IL-33 (mouse) ELISA Kit AG-45A-0038EK-KI01 AG-45A-0038T-KI01 Twinlex 2 x AG-45A-0038-KI01 entalex 5 x SECIES REACTIVITY: Mouse 10 pg/ml 16 to 1000 pg/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Culture Supernatant, lasma, Serum 2 ALICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; IHC: Immunohistochemistry I: Immunoprecipitation; WB: Western blot FORMULATION: F = reservative free SECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; rm = rimate

3 Broad anel of IL-33 Antibodies & roteins ANTIBODIES ID SIZE SOURCE/ISOTYE SECIES ALICATIONS IL-33 (human), mab (IL33305B) AG-20A μg 100 μg Ms IgG2a Hu ELISA, FUNC, IHC (S), I, WB IL-33 (human), pab AG-25A μg Rb Hu ELISA, IHC, WB IL-33 (human), pab (Biotin) AG-25A-0045B 50 μg Rb Hu ELISA, IHC, WB IL-33 (mouse), mab (rec.) (blocking) (Bondy-1-1) AG-27B μg Ms IgG2b Ms ELISA, FUNC IL-33 (mouse), mab (rec.) (blocking) (Bondy-1-1) (preservative free) AG-27B-0013F 100 μg 500 μg 1 mg Ms IgG2b Ms ELISA, FUNC IL-33 (mouse), mab (rec.) (Carly-1-4) AG-27B μg Hu IgG2 Ms ELISA, WB IL-33 (mouse), pab AG-25A μg Rb Ms ELISA, WB IL-33, mab (IL33026B) AG-20A μg 100 μg Ms IgG1 Hu, Ms ELISA, I, WB IL-33, mab (IL33068A) AG-20A μg 100 μg Ms IgG2b Hu ELISA, WB ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN IL-33 (homeodomain-like helix-turn-helix) (human) AG-40A-0162 E. coli Hu N/A (rec.) (His) IL-33 (human) (rec.) AG-40A μg HEK 293 cells Hu 0.1EU/μg IL-33 (human) (rec.) (His) AG-40A μg E. coli Hu 1EU/μg IL-33 (human) (rec.) (His) CHI-HR μg 50 μg E. coli Hu 1EU/μg IL-33 (human) (rec.) (untagged) AG-40B x E. coli Hu 0.1EU/μg IL-33 (human) (rec.) (untagged) (BULK) AG-40B-0038AA 500 μg E. coli Hu 0.1EU/μg IL-33 (mouse) (rec.) AG-40B x E. coli Hu, Ms 0.1EU/μg IL-33 (mouse) (rec.) (BULK) AG-40B-0041AA 500 μg E. coli Hu, Ms 0.1EU/μg IL-33 (mouse) (rec.) (His) AG-40A μg E. coli Ms 1EU/μg IL-33 Receptor ST2 Reagents ANTIBODIES ID SIZE SOURCE/ISOTYE SECIES ALICATIONS ST2 (human), mab (ST33868) AG-20A μg 100 μg Ms IgG1 Hu ELISA, WB ST2 (human), pab AG-25A μg Rb Hu ELISA, WB ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN ST2 (human) (rec.) AG-40A μg HEK 293 cells Hu 0.1EU/μg ST2 (human):fc (human) (rec.) AG-40A μg HEK 293 cells Hu 0.1EU/μg IL-1 Subfamily Members IL-1, IL-1 and IL-1 Receptors ANTIBODIES ID SIZE SOURCE/ISOTYE SECIES ALICATIONS IL-1R2 (mouse), mab (rec.) (raxy-1-1) AG-27B μg Ms IgG2b Ms ELISA, FACS IL-1 (mouse), mab (Bamboo-1) AG-20B μg Ms IgG Ms ELISA, WB IL-1 (mouse), mab (Bamboo-2) AG-20B μg Ms IgG1 Ms ELISA IL-1 (mouse), mab (Bamboo-2) (Biotin) AG-20B-0058B 100 μg Ms IgG1 Ms ELISA ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN IL-1 (human) (rec.) (His) CHI-HR-20001A 50 μg E. coli Hu 0.1EU/μg IL-1R1 (human):fc (human) (rec.) AG-40B μg 3 x 50 μg HEK 293 cells Hu 0.1EU/μg IL-1 (human) (rec.) (His) CHI-HR-20001B 50 μg E. coli Hu 0.1EU/μg IL-1 (human) (rec.) (untagged) AG-40B x E. coli Hu 0.1EU/μg IL-1 (mouse) (rec.) (untagged) AG-40B x E. coli Ms 0.1EU/μg Tools for Inflammasomes Research needed? Browse The Inflammasomes Expert! IL-1 (mouse) ELISA Kit AG-45B-0003-KI01 SECIES REACTIVITY: Mouse 9 pg/ml 16 to 1000 pg/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Culture Supernatant, lasma, Serum For updated prices and additional information visit or contact your local distributor. International Edition 3

4 IL-36 / / [IL-1F6/8/9] roinflammatory Cytokines & IL-36Ra IL-36 exists in three different forms, IL-36, and (IL-1F6, IL-1F8 and IL-1F9, which have identical biological effects. The IL-36 cytokines do not contain a pro-domain, but a recent study suggests that trimming of several amino acids from the N-terminus increases their specific activity dramatically. The IL-36 family members all bind to the same receptor IL-36R (formerly designated IL-1RL2 or IL-1Rrp2), which is expressed by dendritic cells, CD4 + T cells and macrophages. Binding to this receptor in the presence of the co-receptor IL-RAc results in signal transduction via NF- B or MAK, activating similar intracellular signals as IL-1 and inducing the release of proinflammatory cytokines, such as IL-6, IL-8, IL-17, IL-22 and IL-23. IL-36 family members are abundantly expressed in skin and other tissues specific for each member and can be highly expressed in monocytes. Dendritic cells and naïve CD4 + T cells are targets of the different forms of IL-36 leading to a Th1 response. IL-36 cytokines play a major role in skin inflammation and psoriasis as proinflammatory factors. They may represent potential targets for immune-mediated inflammatory conditions or, alternatively, could be used as adjuvants in vaccination. IL-36Ra (IL-1F5) is a highly and specific IL-36 antagonist that binds to IL-36R but does not recruit the co-receptor and consequently fails to transmit the signal. IL-36Ra inhibits the production of proinflammatory cytokines, including IL-12, IL-1, IL-6, TNF- and IL-23. Mutated IL-36Ra with higher activity has been found in patients with pustular psoriasis, leading to enhanced production of inflammatory cytokines (IL-8 in particular) by keratinocytes. SELECTED REVIEWS: IL-36 a new member of the IL-1 family cytokines: D. Tripodi, et al.; J. Biol. Regul. Homeost. Agents. 26, 7 (2012) (Review) IL-36 in psoriasis: J.E. Towne & J.E. Sims; Curr. Opin. harmacol. 12, 486 (2012) CYTOLASM NUCLEUS IL-1RAc IKK IκB p50 p65 Ac IL-36α/β/γ? TIR Domain IL-1Rrp2 Antagonist IL-36Ra SIGGIR/TIR8 Anti-inflammatory Signal roteasomal Degradation A-1 ERK roinflammatory Cytokine & Chemokine Synthesis No Signal MAKKs JNK IL-1Rrp2 FIGURE: IL-36 Signaling athway IL-36Ra (aa 2-155) (human) (rec.) AG-40B x SOURCE: E. coli. SECIES: Human. ENDOTOXIN: 0.05EU/μg. IL-36Ra (aa 3-156) (mouse) (rec.) AG-40B x SOURCE: E. coli. SECIES: Mouse. ENDOTOXIN: 0.05EU/μg. IL-36 (aa 8-160) (mouse) (rec.) AG-40B-0098 SOURCE: E. coli. SECIES: Mouse. ENDOTOXIN: 0.01EU/μg. IL-36 (aa ) (mouse) (rec.) AG-40B-0099 SOURCE: E. coli. SECIES: Mouse. ENDOTOXIN: 0.01EU/μg. IL-36 (aa 5-157) (human) (rec.) AG-40B-0117 SOURCE: E. coli. SECIES: Human. ENDOTOXIN: 0.01EU/μg. IL-36 (aa ) (mouse) (rec.) AG-40B-0100 SOURCE: E. coli. SECIES: Mouse. ENDOTOXIN: 0.01EU/μg. IL-36Ra (human) Matched air Detection Set AG-46B-0006-KI01 For 5 x SECIES REACTIVITY: Human 0.5 ng/ml 0.78 to 50 ng/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Culture Supernatant IL-6 pg/ml IL-36Ra 1ug/ml Unstim IL-36Ra 1ug/ml IL-36α 10ng/ml IL-36α 100ng/ml IL-36β 1ng/ml IL-36β 10ng/ml IL-36β 100ng/ml IL-36γ 1ng/ml IL-36γ 10ng/ml IL-36γ 100ng/ml LS 10ng/ml IL-36Ra/IL-36α 10 IL-36Ra/IL-36α 100 IL-36Ra/IL-36β 1 IL-36Ra/IL-36β 10 IL-36Ra/IL-36β 100 IL-36Ra/IL-36γ 1 IL-36Ra/IL-36γ 10 IL-36Ra/IL-36γ 100 IL-36Ra/LS FIGURE: IL-36Ra (aa 3-156) (mouse) (rec.) (AG-40B-0096) inhibits IL-36, IL-36, IL-36 -dependent expression of IL-6 in mouse BMDCs. METHOD: Bone marrow cells were differentiated into bone marrowderived dendritic cells (BMDCs) using GM-CSF (20ng/ml) for 10 days. BMDCs were cultured with or without IL-36Ra, IL-36, IL-36, IL-36 or LS at indicated concentrations for 72 h. Cell supernatants were then harvested and IL-6 levels determined by ELISA. 4 ALICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; IHC: Immunohistochemistry I: Immunoprecipitation; WB: Western blot FORMULATION: F = reservative free SECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; rm = rimate

5 IL-37 [IL-1F7] An Anti-inflammatory Cytokine IL-37 (IL-1F7) belongs to the IL-1 family. Five splice variants (IL-37a-f) have been identified in humans, however, none of these variants were detected so far in mice. Isoform b is the best characterized and the longest with 218 amino acids and the ability to form homodimers. IL-37 is produced as a propeptide found in the cytoplasm that can translocate to the nucleus following cleavage by caspase-1. The mature form of IL-37b possesses a regulatory role in gene transcription, decreasing the production of LS-stimulated proinflammatory cytokines. Both the precursor and mature IL-37b bind to the receptor IL-18R, but no biological activity (agonistic or antagonistic) has been reported. Recently, it was described that the IL-37b/IL-18R complex uses an accessory protein, such as SIGIRR/TIR8, thereby activating a yet unknown anti-inflammatory pathway. IL-37 has potent anti-inflammatory properties and this function requires SMAD3. When overexpressed in mice, IL-37b decreases production of proinflammatory and Th1/ Th17 cytokines. IL-37 has been detected in various inflammatory diseases and may function as an inhibitor of both innate and adaptive immune responses. It may therefore constitute a treatment target in inflammatory and autoimmune diseases. SELECTED REVIEWS: IL-37 is a fundamental inhibitor of innate immunity: M.F. Nold, et al.; Nat. Immunol. 11, 1014 (2010) Opposing Functions of Classic and Novel IL-1 Family Members in Gut Health and Disease: L.R. Lopetuso, et al.; Front. Immunol. 4, 181 (2013) CYTOLASM Translocation IL-37? Transcription NUCLEUS IL-18Rα No Signal Ac IL-37 IL-18Rβ IL-18Rα? IL-18Rα TIGGIR SIGGIR Anti-inflammatory Signal A-1 Inflammatory Cytokine & Chemokine Synthesis FIGURE: IL-37 Signaling athway? AdipoGen IL-37 ELISA Kits Recommended by Experts IL-37 (human) ELISA Kit AG-45A-0041EK-KI01 AG-45A-0041T-KI01 Twinlex 2 x AG-45A-0041-KI01 entalex 5 x SECIES REACTIVITY: Human 10 pg/ml 16 to 1000 pg/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Culture Supernatant, lasma, Serum IL-37 (human) (IntraCellular) ELISA Kit AG-45A-0040EK-KI01 AG-45A-0040T-KI01 Twinlex 2 x SECIES REACTIVITY: Human 20 pg/ml 31 to 2000 pg/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Lysate IL-18/37 Subfamily Member Antibodies and roteins RODUCT NAME ID SIZE SOURCE SECIES ALICATIONS IL-37 (human), pab AG-25A μg Rb Hu ELISA, IHC, I, WB RODUCT NAME ID SIZE SOURCE SECIES ENDOTOXIN IL-37 (aa ) (human) (rec.) (His) AG-40A μg E. coli Hu 0.1EU/μg IL-37 (human) (rec.) (His) AG-40A μg E. coli Hu N/A IL-18 (human) (rec.) (His) CHI-HR μg E. coli Hu 1EU/μg SIGIRR (human):fc (human) (rec.) AG-40A μg HEK 293 cells Hu 0.1EU/μg For updated prices and additional information visit or contact your local distributor. International Edition 5

6 IL-38 [IL-1F10] A ro- or Anti-inflammatory Cytokine? IL-38 (IL-1F10) is the latest member of the IL-1 family. It is expressed as a 152 aa precursor that is secreted although it lacks a signal peptide. It shares 41% homology with IL-1Ra and 43% with IL-36Ra. Like for all IL-36 family members, trimming of several amino acids from the N-terminus may also occur on IL-38 to increase its activity. Little is known about IL-38. Its receptor is still to be discovered, although a study reported binding of IL-38 to the IL-36 receptor. IL-38 through this binding to the IL-36 receptor may exert inhibitory effects resembling those of IL-36Ra, including inhibition of the production of Th1 cytokines, IL-17 and IL-22. IL-38 (IL-1F10) mrna is expressed in the basal epithelium of skin and by proliferating B cells in tonsil germinal center. It is also found in heart, placenta, fetal liver, spleen and thymus. Its expression in a variety of immune tissues and its similarity to IL-1Ra and IL-36Ra suggest a role of IL-38 in the inflammatory response. IL-38 (human) Matched air Detection Set AG-46A-0002-KI01 For 5 x SECIES REACTIVITY: Human 40 pg/ml 63 to 4000 pg/ml DETECTION TYE: ASSAY TYE: SAMLE TYE: Cell Culture Supernatant SELECTED REVIEWS: New insights in the immunobiology o IL-1 family members: F.L. van de Veerdonk & M.G. Netea; Front. Immunol. 4, newcomers creating new numbers in rheumatology: IL-34 to IL-38: G. Clavel, et al.; Joint Bone Spine (Epub ahead of print) (2013) ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN IL-38 (aa ) (human) (rec.) AG-40B-0129 E. coli Hu 0.01EU/μg IL-38 (aa 3-152) (mouse) (rec.) (FLAG) AG-40B-0101 E. coli Ms 0.01EU/μg IL-38 (human) (rec.) (His) AG-40A μg E. coli Hu 1EU/μg IL-6 (mouse) otent Hybridoma Growth Factor No need to add expensive supplement media to your hybridoma cell culture. Interleukin-6 (IL-6; B cell hybridoma growth factor) is a pluripotent cytokine which plays a role in the pathophysiology of severe infection, in immune regulation, hematopoiesis, inflammation, oncogenesis and acute phase reaction. IL-6 is produced by activated T cells, monocytes or macrophages, endothelial cells and fibroblasts. IL-6 is also a myokine, which functions in energy metabolism. It stimulates the growth and survivability of certain B cells and T cells and, when added in media for hybridoma growth, it eliminates the need for feeder cells. It is the key component of many growth culture medias. IL-6 binds to a unique receptor system comprising of IL-6R, a receptor specific for IL-6 and gp130, the common signal transducer of cytokines related to IL-6. Signal transduction through gp130 is mediated by two pathways: the JAK-STAT (Janus family tyrosine kinase-signal transducer and activator of transcription) pathway and the Ras-MAK (mitogen-activated protein kinase) pathway. Abnormal overproduction of IL-6 is responsible for the pathogenesis of various inflammatory diseases, such as plasmacytoma/myeloma and several chronic inflammatory proliferative diseases. ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN IL-6 (mouse):fc (human) (rec.) AG-40B x HEK 293 cells Ms 0.01EU/μg IL-6 (mouse):fc (mouse) (rec.) (non-lytic) CHI-MF μg 3 x 50 μg CHO cells Ms 0.06EU/μg For a Full anel of roducts visit 6 ALICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; IHC: Immunohistochemistry I: Immunoprecipitation; WB: Western blot FORMULATION: F = reservative free SECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; rm = rimate

7 Other Interleukin Antibodies & roteins ANTIBODIES ID SIZE SOURCE/ISOTYE SECIES ALICATIONS IL-2 (human), mab (ANC7F7) ANC μg Ms IgG1 Hu FACS IL-2RA [CD25] (human), mab (7G7B6) ANC μg Ms IgG2a Hu FACS, IHC IL-2RB [CD122] (human), mab (9A2) ANC μg Ms IgG2a Hu FACS, FUNC IL-7RA [CD127] (human), mab (ANC8F2) ANC μg Ms IgG1 Hu FACS IL-17A (human), mab (17-142F) AG-20A μg 100 μg Ms IgG1 Hu ELISA, FACS ROTEINS IL-2 (human) (rec.) (His) IL-2 (human):fc (human) (rec.) IL-2 (human):fc (human) (rec.) (non-lytic) IL-2 (mouse):fc (mouse) (rec.) IL-2 (mouse):fc (mouse) (rec.) (non-lytic) IL-2 (rat) (rec.) (His) IL-2 Superkine (Fc) IL-4 (human) (rec.) (His) IL-4 (human):fc (human) (rec.) IL-4 (human):fc (human) (rec.) (non-lytic) IL-4 (mouse) (rec.) (His) IL-4 (mouse):fc (mouse) (rec.) (non-lytic) IL-6 (human) (rec.) (His) IL-6 (human):fc (human) (rec.) IL-6 (human):fc (human) (rec.) (non-lytic) IL-6R (human):fc (human) (rec.) IL-7RA [CD127] (human)-muig Fusion rotein IL-8 (human) (rec.) (His) IL-8 (human):fc (human) (rec.) (non-lytic) ID CHI-HR CHI-HF CHI-HF CHI-MF CHI-MF CHI-RR AG-40B-0111 CHI-HR CHI-HF CHI-HF CHI-MR CHI-MF CHI-HR CHI-HF CHI-HF CHI-HF-21006R ANC CHI-HR CHI-HF ROTEINS IL-10 (human) (rec.) (His) IL-10 (human):fc (human) (rec.) IL-10 (human):fc (human) (rec.) (non-lytic) IL-10 (mouse):fc (mouse) (rec.) (non-lytic) IL-15 (mutant) (human):fc (human) (rec.) IL-16 (human) (rec.) (His) IL-17A (human):fc (human) (rec.) IL-17A (mouse) (rec.) (His) IL-17F (human) (rec.) (His) IL-21 (human):fc (human) (rec.) (non-lytic) IL-21 (mouse):fc (mouse) (rec.) (non-lytic) IL-21 (mutant) (human) (rec.) (His) IL-21R (human):fc (human) (rec.) IL-21R (mouse):fc (mouse) (rec.) (non-lytic) IL-22 (human):fc (human) (rec.) IL-22 (human):fc (human) (rec.) (non-lytic) IL-22 (mouse) (rec.) (His) IL-22 (mouse):fc (mouse) (rec.) IL-22 (mouse):fc (mouse) (rec.) (non-lytic) ID CHI-HR CHI-HF CHI-HF CHI-MF CHI-HF-21015M CHI-HR AG-40A-0066 CHI-MR-10017A CHI-HR-20017F CHI-HF CHI-MF CHI-HR-20021M CHI-HF-21021R CHI-MF-12021R CHI-HF CHI-HF CHI-MR CHI-MF CHI-MF IL-2 Superkine (Fc) Highly Active & Low Toxic Artificial rotein AG-40B-0111-C010 AG-40B Multiack 3 x SOURCE: HEK 293 cells. SEQUENCE: The extracellular domain of human IL-2 (aa ) (mutant H9 containing the mutations L80F / R81D / L85V / I 86V / I92F) is fused at the C-terminus to the Fc portion of human IgG2. SECIFICITY: Binds to human and mouse IL-2R. BIOLOGICAL ACTIVITY: Triggers far greater antitumor responses than native IL-2 in vivo but with lower toxicity. IL-2 Superkine (Fc) is an artificial variant of IL-2 containing mutations at positions L80F / R81D / L85V / I 86V / I92F. These mutations are located in the molecule s core that acts to stabilize the structure and to give it a receptor-binding conformation mimicking native IL-2 bound to CD25. It effectively eliminates the functional requirement of IL-2 for CD25 expression and directly triggers proliferation of T cells. LIT: Exploiting a natural conformational switch to engineer an interleukin-2 superkine : AM. Levin, et al.; Nature 484, 529 (2012) OD 450nm IL-2 Superkine (Fc) IL-2 (human):fc (human) ng/ml FIGURE: Binding of IL-2 Superkine (Fc) (AG-40B-0111) to IL-2R (human) is increased 10 fold compared to IL-2 (human):fc (human). For updated prices and additional information visit or contact your local distributor. International Edition 7

8 IL-35 An Inhibitory Cytokine & The IL-12 Family HIGHLIGHT IL-35 is a newly discovered heterodimeric cytokine. It is a member of the IL-12 family, which belongs to the IL-6 superfamily and includes IL-12, IL-23, IL-27 and IL-35. These cytokines are heterodimeric proteins and are made up of an -chain (p19, p28 or p35) and a -chain (p40 or Ebi3) (see Figure). IL-12 signals through IL-12R 1 and IL-12R 2; IL-23 signals via IL-12R 1 and IL-23R; IL-27 uses gp130 and IL-27R and IL-35 signals with gp130 and IL-12R 2 or through homodimers of each chain. Downstream effects are carried out by various members of the JAK-STAT family. IL-12 is critical for responses to pathogens and induces IFN-. IL-23 has a proinflammatory function in response to pathogens and also stabilizes Th17 cells. IL-27 is mostly inhibitory and is secreted by local ACs to limit inflammatory responses. IL-35 suppresses T cell proliferation and converts naïve T cells into IL-35-producing induced regulatory T cells (itr35 cells, a type of T regulatory cells that do not express Foxp3). itr35 cells can mediate self tolerance and prevent autoimmunity in an IL-35-dependent manner. Effects of IL-35 s inhibitory role may be mediated by IL-10. IL-35 is considered to have a potential therapeutic effect against immune diseases and could promote the development of different kinds of vaccines for immunotherapy against cancer and be promising to cure autoimmune and inflammatory diseases. roinflammatory IL-12Rβ1 IL-23 IL-12 IL-27 IL-35 p19 p35 p28 p35 p40 p40 EBI3 EBI3 IL-23R IL-12Rβ1 IL-12Rβ2 JAK2 JAK2 JAK1 TYK2 TYK2 JAK2 JAK1 JAK2 STAT3 STAT3 gp130 CYTOLASM NUCLEUS IL-27R (WSX-1) STAT1 STAT3 STAT1 STAT3 gp130 IL-12Rβ2 STAT1 STAT1 Inhibitory SELECTED REVIEWS: IL-12 family cytokines: immunological playmakers: D.A. Vignali & V.K. Kuchroo; Nat. Immunol. 13, 722 (2012) Interleukin-35: the future of hyperimmunerelated diseases? S. Ye, et al.; J. Interferon Cytokine Res. 33, 285 (2013) rotumor Immune & roinflammatory Responses Antitumor & roinflammatory Responses / Cytokine roduction Immune Suppression FIGURE: IL-12 Family Signaling athway ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN IL-35 (human):fc (human) (rec.) CHI-HF μg CHO cells Hu 0.06EU/μg IL-35 (mouse):fc (human) (rec.) CHI-MF μg CHO cells Ms 0.06EU/μg ANTIBODIES ID SIZE SOURCE SECIES ALICATIONS IL-23p19 (human), mab (I 178G) AG-20A μg 100 μg Rat IgG2a Hu ELISA, WB IL-23p19 (human), mab (I 338H) AG-20A μg 100 μg Rat IgG2a Hu ELISA, WB IL-23p19, pab AG-25A μg Rb Hu, Ms ELISA, WB IL-23p40 (human), mab (I 1108) AG-20A μg 100 μg Ms IgG1 Hu ELISA, WB ROTEINS ID SIZE SOURCE SECIES ENDOTOXIN IL-12 (mouse):fc (human) (rec.) CHI-MF μg CHO cells Ms 0.06EU/μg IL-23 (human) (rec.) AG-40A μg HEK 293 cells Hu 0.1EU/μg IL-23 (mouse):fc (human) (rec.) CHI-MF μg CHO cells Ms 0.06EU/μg IL-27 (human):fc (human) (rec.) CHI-HF μg CHO cells Hu 0.06EU/μg IL-27 (human):fc (human) (rec.) (non-lytic) CHI-HF μg CHO cells Hu 0.06EU/μg IL-27 (mouse):fc (human) (rec.) CHI-MF μg CHO cells Ms 0.06EU/μg Epstein-Barr Virus-induced Gene 3 (human):fc (human) (rec.) AG-40A μg HEK 293 cells Hu 0.1EU/μg EUROE/REST OF WORLD Adipogen International TEL FAX info@adipogen.com For local distributors please visit our website. NORTH & SOUTH AMERICA Adipogen Corp. TEL FAX info-us@adipogen.com SOUTH KOREA/ASIA Adipogen, Inc. TEL FAX info-kr@adipogen.com OCT 2013