TNF Superfamily Linking Autoimmune Diseases, TNF Inflammation and Cancer

International Edition

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E TNF Superfamily Linking Autoimmune Diseases, TNF Inflammation and Cancer

THE Adipogen's Team has 15 years of experience in manufacturing TNF superfamily reagents. This expertise has been acquired by an established long-term collaboration with renowned labs of key players in the field including Professor Jürg Tschopp, who first cloned the TNF superfamily ligands BAFF and APRIL. The tumor necrosis factor (TNF) superfamily (TNFSF) is composed of 19 structurally related proteins (ligands) that bind to one or more molecules from the TNF receptor superfamily (TNFRSF), a family of 30 structurally similar receptors. The TNFSF ligands are either membrane-anchored or soluble trimers that cluster their cognate cell surface receptors to initiate signal transduction. The unique structural features of the TNFSF ligands and receptors link these molecules to cell growth, cell survival or cell death, although some molecules can activate both inflammatory and cell death pathways, dependent on target cell types and other extrinsic stimuli. Many of the TNFRSF molecules are expressed in cells of the immune system, which implies that they may be central to autoimmune and inflammatory diseases as well as cancer. However, their function is not restricted to immune cells, members of these superfamilies have been linked to an array of pathophysiologies, including cancer, neurologic diseases, cardiovascular diseases, pulmonary diseases, autoimmune diseases and metabolic diseases. TNFSF and TNFRSF genes have been found to display polymorphisms that are linked to human diseases, which implies that interventions targeting these molecules may be efficacious in treating several health-related problems. Therefore, it is not surprising that several biologics targeting TNFSF and TNFRSF molecules are in ongoing clinical trials for autoimmune and inflammatory diseases and cancer.

Membrane Membrane TNFSF Soluble TNFSF TNFSF

TNFRSF TNFRSF

TRADD or FADD TRAFs BAX or BID IKKα, IKKβ, IKKγ IKKα, IKKβ, IKKγ NIK Pro-caspase 8 Pro-caspase 10

Mitochondria IKKα IκBα Caspase-8 MAPK AKT Cytochrome c p50 RELA p100 RELB Pro-caspase-9 Caspase-10

Induction of cell cycle proteins, anti-apoptotic proteins, Induction of apoptosis via activation of inflammatory cytokines and chemokines caspase-3, caspase-6 and caspase-7

FIGURE: Pro-inflammatory and death-inducing activity of TNFSF members. Adapted from M. Croft, et al.; Nat. Rev. Drug Disc. 12, 147 (2013) LIGAND: RECEPTOR: CD40L [TNFSF5] CD40 [TNFRSF5]

CD40L (CD154) binds to CD40, which is essential in mediating event in Alzheimer disease pathogenesis. CD40 is constitutive- a broad variety of immune and inflammatory responses in- ly expressed by antigen presenting cells, including dendritic cluding T cell-dependent immunoglobulin class switching, cells, B cells and macrophages. Consistent with its widespread memory B cell development and germinal center formation. expression on normal cells, CD40 is also expressed on a wide The interaction of CD40-CD40L is necessary for amyloid--in- range of tumor cells. CD40L levels are increased in serum and duced microglial activation, and thus is thought to be an early in inflamed tissues of patients with autoimmune diseases.

MegaLigands

MegaLigands are high activity constructs in which two trimeric TNFSF ligands are artificially linked via the oligomeric collagen domain of ACRP30. MegaCD40L is a potent B cell activator. MegaCD40L very effectively mimics the natural membrane-assisted aggregation of CD40L. It is the most potent alternative to activate CD40 with the advantage that no oligomerization reagent (enhancer) is required.

BULK available! MegaCD40L

MegaCD40L, Soluble (human) (rec.) cells 80 MegaCD40L (h) AG-40B-0010-C010 10 μg 70 AG-40B-0010-3010 MultiPack 3 x 10 μg CD40L (h) 60 CD40L (h) + enhancer

SOURCE: CHO cells. SEQUENCE: Human CD40L (aa 116-261) is 50 fused at the N-terminus to mouse ACRP30headless (aa 18- 40 111) and a FLAG®-tag. SPECIFICITY: Binds to human CD40. BIOLOGICAL ACTIVITY: Induces B cells activation (as demon- 30 20 strated by dose-dependent upregulation of CD86) (ED50: Percentage /Cd19 posive of CD86 4 0

37 12 ng /ml

ENDOTOXIN CONTENT: 333 111

<1ng/ml). <0.01EU/μg purified protein 1,37 0,46 0,15 0,05 0,02 1000 (LAL test; Lonza). FIGURE: MegaCD40L, Soluble (human) (rec.) (Prod. No. AG-40B-0010) LITERATURE REFERENCES: does not need an enhancer to induce B cells activation. IgG subclass switch capacity is low in switched and in IgM-only, but high in METHOD: IgD+IgM+, post-germinal center (CD27+) human B cells: C. Werner-Favre, PBL cells were incubated in 96-well plates (2x105 cells/well in et al.; Eur. J. Immunol. 31, 243 (2001) 100μl RPMI supplemented with 10% FCS) for 24 hours at 37°C with the indicated concentration of MegaCD40L, Soluble (human) (rec.) or CD40L Two adjacent trimeric Fas ligands are required for Fas signaling and (human) in the presence and absence of 1μg/ml Enhancer (Prod. No formation of a death-inducing signaling complex: N. Holler, et al.; Mol. AG-35B-0001). Cells were washed with PBS and stained with 2μl each Cell. Biol. 23, 1428 (2003) CD86-PE and CD19-FITC in 50μl FACS buffer (PBS, 5% fetal calf serum, Impaired CD40L signaling is a cause of defective IL-12 and TNF-alpha 0.02% azide) for 20 min. at 4°C in the dark. After two washes in FACS production in Sezary syndrome: circumvention by hexameric soluble buffer, samples were then analyzed by flow cytometry. CD40L: L.E. French, et al.; Blood 105, 219 (2005)

Only available from AdipoGen®

www.adipogen.com MegaCD40L, Soluble (mouse) (rec.) MegaCD40L, Soluble (rat) (rec.) AG-40B-0020-C010 10 μg AG-40B-0107-C010 10 μg AG-40B-0020-3010 MultiPack 3 x 10 μg AG-40B-0107-3010 MultiPack 3 x 10 μg BULK available for in vivo studies! Bulk available for in vivo studies!

APPLICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; FORMULATION: PF = Preservative free 2 IHC: Immunohistochemistry IP: Immunoprecipitation; WB: Western blot SPECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; Prm = Primate BULK available! THE STANDARD Mouse CD40 Antibody for in vivo studies Activates B and NK cells in vivo and in vitro

PRODUCT NAME PID SIZE ISOTYPE APPLICATION SPECIES CD40 (mouse), mAb (FGK45) AG-20B-0036 100 μg | 500 μg Rt IgG2a FACS, FUNC (Activation) Ms CD40 (mouse), mAb (FGK45) (PF) AG-20B-0036PF 100 μg | 500 μg Rt IgG2a FACS, FUNC (Activation) Ms www.adipogen.com

Human CD40/CD40L Proteins & Antibodies

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES CD40 (human):Fc (human) (rec.) AG-40B-0083 50 μg | 3 x 50 μg HEK 293 cells <0.1EU/μg Hu, Ms ANTIBODIES PID SIZE ISOTYPE APPLICATION SPECIES CD40 (human), mAb (BE-1) ANC-189-020 100 μg Ms IgG1 ELISA, FACS, FUNC, IP Hu CD40L [CD154] (human), mAb (24-31) ANC-353-020 100 μg Ms IgG1 ELISA, FACS, FUNC Hu, Prm (Blocking), IHC, WB

LIGANDS: RECEPTORS: LT [TNFSF3] LTR [TNFRSF3] LIGHT [TNFSF14] HVEM [TNFRSF14] DcR3 [TNFRSF6B]

LT (LT12) binds to the LTR activating two different NF- noma HT-29. It also binds to decoy receptor 3 (DcR3) and HVEM. B pathways that lead to distinct patterns of gene induction, B and T lymphocyte associated (BTLA; CD272) is an Ig domain including selected chemokines and the cytokine BAFF, which superfamily member. The herpes virus entry mediator (HVEM; is essential for the survival of mature B lymphocytes. LTR CD270) can act as a molecular switch that modulates T cell activates the classical NF-B (relA/p50) pathway, like the type activation by propagating positive signals from the TNF-re- 1 TNF receptor (TNF-R1), that regulates proinflammatory genes. lated ligand LIGHT, or inhibitory signals through the Ig su- However, LTR, unlike TNF-R1, also activates the processing perfamily member BTLA. The binding site on HVEM for BTLA of p100 to form RelB/p52 complexes, which activate genes is conserved in the orphan TNF receptor UL144, present in involved in lymphoid organ formation and lymphocyte survival. human cytomegalovirus (CMV). UL144 binds BTLA, but not LIGHT (CD258) binds to LTR. It activates NF-B, stimulates the LIGHT, and inhibits T cell proliferation, selectively mimicking proliferation of T cells and inhibits growth of the adenocarci- the inhibitory cosignaling function of HVEM.

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES BTLA (human)-muIg Fusion Protein ANC-542-020 25 μg CHO cells N/A Hu HVEM (human)-muIg Fusion Protein ANC-531-020 25 μg CHO cells N/A Hu LIGHT, Soluble (human) (rec.) AG-40B-0009 10 μg | 3 x 10 μg CHO cells <0.01EU/μg Hu, Ms LTR (human):Fc (human) (rec.) (non-lytic) CHI-HF-220LTBR 100 μg CHO cells <0.06EU/μg Hu ANTIBODIES PID SIZE ISOTYPE APPLICATION SPECIES BTLA (human), mAb (6F4) AG-20B-0049 100 μg Rt IgG1 ELISA, FACS, Hu FUNC (Inhibition) HVEM (human), mAb (ANC3B7) ANC-270-020 100 μg Ms IgG2a ELISA, FACS Hu LTR (mouse), mAb (3C8) AG-20B-0041 100 μg Rt IgG1 FUNC (Activation) Ms LTR (mouse), mAb (3C8) (PF) AG-20B-0041PF 100 μg Rt IgG1 FUNC (Activation) Ms LTR (mouse), mAb (4H8 WH2) AG-20B-0008 100 μg Rt IgG2a FACS, Ms FUNC (Activation) LTR (human), mAb (ANCLTR2/9E2) ANC-267-020 100 μg Ms IgG1 ELISA, FACS Hu

Visit www.adipogen.com for additional preservative free and labeled antibodies!

For updated prices and additional information visit www.adipogen.com or contact your local distributor. 3 LIGAND: RECEPTORS: FasL [TNFSF6] Fas [TNFRSF6] DcR3 [TNFRSF6B]

FasL (CD95L; CD178) binds to Fas (CD95), a receptor that trans- mediated apoptosis. Stimulation of Fas has also been reported duces the apoptotic signal into cells. It is involved in cytotoxic to trigger non-apoptotic pathways. It has been shown that T cell-mediated apoptosis and in T cell development. The membrane-bound FasL is essential for the cytotoxic activity, formation of the Fas death-inducing signaling complex (DISC) whereas soluble FasL appears to promote autoimmunity and is the initial step of Fas signaling. Activation of procaspase-8 tumorigenesis via induction of non-apoptotic pathways, in at the DISC leads to the induction of death receptor (DR)- particular NF-B. FasL binds also to decoy receptor 3 (DcR3).

THE STANDARD Fas Antibody Widely cited antibody for in vivo application! Induces apoptosis.

PRODUCT NAME PID SIZE ISOTYPE APPLICATION SPECIES Fas (human), mAb (APO-1-3) (PF) AG-20B-0062PF 50 μg Ms IgG3 FACS, FUNC (Activation), IP, WB Hu

BULK available! NEW MegaFasL

OD 2,5 MegaFasL, Soluble (human) (rec.) AG-40B-0130-C010 10 μg 2

AG-40B-0130-3010 MultiPack 3 x 10 μg 1,5 MegaFasL (h) Fc:FasL (h) 1 SOURCE: SEQUENCE: FasL (h) HEK 293 cells. Human FasL (aa 139-281) FasL (h) + enhancer is fused at the N-terminus to mouse ACRP30headless 0,5 (aa 18-111) and a FLAG®-tag. SPECIFICITY: Binds to human 0

and mouse Fas. BIOLOGICAL ACTIVITY: Induces apoptosis of 7,81 3,91 1,95 0,98 0,49 0,24 0,12 0,06 0,03 0,02 0,01 62,50 31,25 15,63 500,00 250,00 125,00 ng /ml human Jurkat T cells at a concentration of <1ng/ml. ENDO- 1.000,00 TOXIN CONTENT: <0.02EU/μg purified protein (LAL test; Lonza). FIGURE: Oligomerisation of FasL efficiently triggers Jurkat cell death. LITERATURE REFERENCES: METHOD: Jurkat cells were treated O/N with the indicated concentrations Two adjacent trimeric Fas ligands are required for Fas signaling and of several FasL proteins (2 fold-dilutions, first concentration of 1000ng/ml). formation of a death-inducing signaling complex: N. Holler, et al.; Mol. Cell death was quantified using PMS/MTS. The oligomeric FasL recombinant Cell. Biol. 23, 1428 (2003) proteins (MegaFasL, Soluble (human), Fc:FasL, Soluble (human) and FasL, Soluble (human) + Enhancer) kill Jurkat cells at IC <0.2ng/ml. A Fas agonist induces high levels of apoptosis in haematological 50 malignancies: P. Greaney, et al.; Leuk. Res. 30, 415 (2006)

Also available

ANTIBODY PID SIZE ISOTYPE APPLICATION SPECIES FasL (human), mAb (ALF-2.1A) ANC-399-020 100 μg Ms IgG1 ELISA, FACS, Hu FUNC (Blocking) PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES www.adipogen.com Fas (human):Fc (human) (rec.) AG-40B-0082 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms FasL, Soluble (human) (rec.) AG-40B-0001 10 μg | 3 x 10 μg HEK 293 cells <0.05EU/μg Hu, Ms FasL, Soluble (human) (rec.) (BULK) AG-40B-0001AA 500 μg HEK 293 cells <0.05EU/μg Hu, Ms NEW Fc (human):FasL, Soluble (human) (rec.) AG-40B-0132 10 μg | 3 x 10 μg HEK 293 cells <0.01EU/μg Hu, Ms EnhancedFasL, Soluble (human) (rec.) Pack AG-44B-0001 1 Set HEK 293 cells <0.05EU/μg Hu, Ms

APPLICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; FORMULATION: PF = Preservative free 4 IHC: Immunohistochemistry IP: Immunoprecipitation; WB: Western blot SPECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; Prm = Primate LIGAND: RECEPTOR: OX40L [TNFSF4] OX40 [TNFRSF4]

OX40L (CD134L; CD252) acts as a costimulator through OX40:Fc was shown to prevent OX40L from reaching the T cell the interaction with OX40 (CD134) on T cells, stimulating receptors, thus reducing the T cell response. Experiments T cell activation, proliferation and cytokine production. It in mice have demonstrated that OX40:Fc can reduce the is expressed on antigen presenting cells including B cells, symptoms associated with the cytokine storm (an immune www.adipogen.com dendritic cells, mast cells and endothelium. OX40 has a overreaction) while allowing the immune system to fight off critical role in the maintenance of an immune response. viruses successfully.

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES OX40 (human):Fc (human) (rec.) AG-40B-0014 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms OX40 (human) (rec.) (His) CHI-HR-200CD134 25 μg E. coli <0.1EU/μg Hu MegaOX40L, Soluble (mouse) (rec.) AG-40B-0029 10 μg HEK 293 cells <0.01EU/μg Ms, Hu ANTIBODIES PID SIZE ISOTYPE APPLICATION SPECIES OX40 (human), mAb (BerAct35) ANC-355-020 100 μg Ms IgG1 ELISA, FACS, IHC Hu OX40L (human), mAb (ANC10G1) ANC-400-020 100 μg Ms IgG1 ELISA, FACS, FUNC Hu (Blocking)

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LIGAND: RECEPTOR: CD137L [TNFSF9] CD137 [TNFRSF9]

CD137L (4-1BBL) is expressed on activated macrophages, anti-viral immunity, diabetes, in T and B cell co-stimulation dendritic cells and mature B cells. The interaction with its and modulation of the immune response. Crosslinking of receptor CD137 (4-1BB) induces recruitment of TNF receptor- CD137 enhances T cell proliferation, IL-2 secretion survival associated factor 1 (TRAF1) and -2 (TRAF2) and interaction and cytolytic activity. CD137 was shown to enhance immune with the kinase p56lck. CD137 (4-1BB) and CD137L have activity to eliminate tumors in mice. been reported to be involved in tumor rejection, apoptosis,

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES CD137 (human) (rec.) (His) CHI-HR-200CD137 25 μg E. coli <0.1EU/μg Hu CD137 (human):Fc (human) (rec.) AG-40B-0060 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu CD137 (mouse):Fc (human) (rec.) AG-40A-0025 50 μg HEK 293 cells <0.1EU/μg Ms CD137L, Soluble (human) (rec.) AG-40A-0198 50 μg HEK 293 cells <0.1EU/μg Hu CD137L, Soluble (mouse) (rec.) AG-40A-0020 50 μg HEK 293 cells <0.1EU/μg Ms CD137L, Soluble (mouse) (rec.) (His) AG-40A-0067 10 μg | 50 μg E. coli <1EU/μg Ms ANTIBODIES PID SIZE SOURCE/ISOTYPE APPLICATION SPECIES CD137 (human), pAb AG-25A-0018 100 μg Rb FACS, WB Hu CD137 (mouse), mAb (M4173) AG-20A-0072 50 μg | 100 μg Rt IgG1 FACS, WB Ms CD137 (mouse), mAb (M4202) AG-20A-0062 50 μg Rt IgG1 FACS Ms CD137 (mouse), pAb AG-25A-0085 100 μg Rt FACS, WB Ms CD137L (human), mAb (41B436) AG-20A-0031 50 μg | 100 μg Ms IgG1 ELISA, FACS, WB Hu CD137L (human), mAb (41B446) AG-20A-0063 50 μg Ms IgG1 FACS, WB Hu

Ask for BULK quantities!

For updated prices and additional information visit www.adipogen.com or contact your local distributor. 5 LIGAND: RECEPTORS: TRAIL [TNFSF10] TRAIL-R1 [TNFRSF10A] TRAIL-R2 [TNFRSF10B] TRAIL-R3 [TNFRSF10C] TRAIL-R4 [TNFRSF10D] OPG [TNFRSF11B]

TNF-related apoptosis-inducing ligand (TRAIL; Apo2L; CD253) (DISC). Aside its apoptotic effect and its importance for cancer is a type II transmembrane protein of about 34kDa. Active regulation, TRAIL and TRAIL-Rs seem to be involved in different TRAIL forms trimers and specifically binds to five distinct pathways and regulatory functions such as non-apoptotic, receptors: TRAIL-R1 (DR4; CD261), -R2 (DR5; CD262), -R3 mitogenic and prosurvival pathways including the MAPKs, the (DcR1; CD263), -R4 (DcR2; CD264) and osteoprotegerin (OPG). protein kinase B (PKB/Akt) and the NF-B signaling cascades. Trimerized TRAIL triggers apoptosis upon ligation of cell They were shown to be involved in bone turnover regulation surface TRAIL-R1 and/or TRAIL-R2 by inducing the formation of and angiogenesis. the so-called multiprotein death-inducing signaling complex

Highly Specific Antibodies for FACS and IHC

PRODUCT NAME PID SIZE ISOTYPE APPLICATION SPECIES TRAIL (human), mAb (HS501) AG-20B-0026 100 μg Ms IgG1 WB Hu TRAIL-R1 (human), mAb (HS101) AG-20B-0022 100 μg Ms IgG1 FACS, ICC, FUNC Hu (Blocking), IP TRAIL-R1 (human), mAb (TR1.02) AG-20B-0027 100 μg Ms IgG2b FACS, IHC, WB Hu TRAIL-R2 (human), mAb (HS201) AG-20B-0023 100 μg Ms IgG1 FACS, ICC, FUNC Hu (Blocking), IP TRAIL-R2 (human), mAb (TR2.21) AG-20B-0028 100 μg Ms IgG1 FACS, IHC, WB Hu TRAIL-R3 (human), mAb (HS301) AG-20B-0024 100 μg Ms IgG1 FACS, ICC Hu TRAIL-R3 (human), mAb (TR3.06) AG-20B-0029 100 μg Ms IgG1 FACS, IHC, WB Hu TRAIL-R4 (human), mAb (HS402) AG-20B-0025 100 μg Ms IgG1 FACS, ICC, IHC, IP Hu TRAIL-R1 to -R4 Flow Cytometry Pack AG-44B-0004 1 Set Ms IgG1 FACS Hu

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Flow Cytometry (FACS) Immunohistochemistry (IHC)

FACS Analysis

FIGURE: Immunohisto- chemistry detection of endo- genous TRAIL-R1, TRAIL-R2 and TRAIL-R3 in paraffin-em- bedded human carcinoma tissues (colon, tonsil) using mAb to TRAIL-R1 (TR1.02) (Prod. No. AG-20B-0027), mAb to TRAIL-R2 (TR2.21) (Prod. No. AG-20B-0028) and mAb to TRAIL-R3 (TR3.06) (Prod. No. AG-20B-0029). www.adipogen.com FIGURE: FACS analysis of surface expression of TRAIL-R1 to TRAIL-R4 with (solid bold line) and without (filled line) 5-FU (100μg/ml) treatment for 16h, compared to control (dashed line) using TRAIL-R1 mAb (HS101) (Prod. No. AG-20B-0022), TRAIL-R2 mAb (HS201) (Prod. No. AG-20B-0023), TRAIL-R3 mAb (HS301) (Prod. No. AG-20B-0024) and For more information see our Brochure TRAIL-R4 mAb (HS402) (Prod. No. AG-20B-0025). "TRAIL & TRAIL Receptors: Effectors of Apoptotic Signaling"

APPLICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; FORMULATION: PF = Preservative free 6 IHC: Immunohistochemistry IP: Immunoprecipitation; WB: Western blot SPECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; Prm = Primate KillerTRAIL and SuperKillerTRAIL Oligomerized TRAIL proteins that do not require a cross-linking enhancer for their potent biological activity. www.adipogen.com

FIGURE: Apoptosis of TRAIL-sensitive cells. Concentration dependence of apoptosis FIGURE: Apoptosis of TRAIL-sensitive cells. Apoptosis induction in BJAB cells by induction in Jurkat and BJAB cells by KillerTRAIL™, Soluble (human) (rec.) (Prod. No. AG- SuperKillerTRAIL™, Soluble (human) (rec.) (Prod. No. AG-40T-0002) reveals killing activity 40T-0001) reveals high activity even at concentrations of 10-100ng/ml. at concentrations of 10-15ng/ml on Jurkat cells even as low as 1-10ng/ml.

izTRAIL Oligomerized TRAIL protein non-toxic to hepatocytes, suitable for in vitro and in vivo use. izTRAIL is a highly active recombinant form of soluble human TRAIL. Due to a trimerizing N-terminal isoleucine zipper (iz) motif the intrinsic trimerization of TRAIL, required for apoptosis-inducing activity of TRAIL, is enhanced when compared to non-tagged soluble human TRAIL (shTRAIL). Therefore, izTRAIL is a potent inducer of apoptosis in many human cancer cells, but not normal human hepatocytes. In addition, the half-life of izTRAIL is about eight-fold higher than the half-life of shTRAIL. These properties render izTRAIL highly suitable for both, in vitro and in vivo use, particularly for studies in which investigators plan to transfer their in vitro results into an in vivo system with human cancer cells in xenotransplant settings FIGURE: Apoptosis of TRAIL-sensitive tumor cells. Concentration dependence of apoptosis examining susceptibility to TRAIL-induced apoptosis. induction in BJAB cells, BL60-cells, U937-cells, and JA3-cells by izTRAIL, Soluble (human) (rec.) (Prod. No. AG-40B-0069) reveals high activity even at concentrations of 10-100ng/ml.

Highly Potent TRAIL Proteins BULK available

PRODUCT NAME PID SIZE SOURCE ENDOTOXIN SPECIES TRAIL, Soluble (human) (rec.) AG-40B-0003 10 μg | 5 x 10 μg E. coli <0.01EU/μg Hu, Ms TRAIL, Soluble (human) (rec.) (BULK) AG-40B-0003AA 500 μg E. coli <0.01EU/μg Hu, Ms EnhancedTRAIL, Soluble (human) (rec.) Pack AG-44B-0002 1 Set E. coli <0.01EU/μg Hu, Ms izTRAIL, Soluble (human) (rec.) AG-40B-0069 10 μg | 5 x 10 μg E. coli <0.1EU/μg Hu KillerTRAIL™, Soluble (human) (rec.) AG-40T-0001 50 μg | 3 x 50 μg E. coli <0.01EU/μg Hu, Ms KillerTRAIL™ (R1 specific), Soluble (human) (rec.) AG-40T-0003 50 μg E. coli <0.1EU/μg Hu SuperKillerTRAIL™, Soluble (human) (rec.) AG-40T-0002 20 μg | 3 x 20 μg E. coli <0.01EU/μg Hu SuperKillerTRAIL™, Soluble (mouse) (rec.) AG-40T-0004 20 μg | 3 x 20 μg E. coli <0.01EU/μg Ms, (Hu) KillerTRAIL™ Dilution & Storage Buffer AG-10T-0001 500 μl <0.1EU/μg TRAIL-R1 (human):Fc (human) (rec.) AG-40B-0070 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms TRAIL-R2 (human):Fc (human) (rec.) AG-40B-0071 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms dcTRAIL-R2 (mouse):Fc (human) (rec.) AG-40B-0094 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms

For updated prices and additional information visit www.adipogen.com or contact your local distributor. 7 LIGAND: RECEPTORS: BAFF [TNFSF13B] BAFF-R [TNFRSF13C] TACI [TNFRSF13B] BCMA [TNFRSF17]

BAFF (BLyS; THANK; CD257) is mainly produced by innate together with IL-6, promotes Ig class-switching and plasma immune cells such as neutrophils, monocytes, macrophages, cell differentiation. Besides its major role in B cell biology, dendritic cells, follicular dendritic cells. T cells, activated B BAFF co-stimulates activated T cells. Deregulated expression cells, some malignant B cells and also non-lymphoid cells like of BAFF leads to autoimmune disorders in mice. In humans, astrocytes, synoviocytes and epithelial cells can also produce elevated levels of soluble BAFF have been detected in the BAFF. BAFF binds three distinct receptors (BAFF-R (CD268), serum of patients with various autoimmune diseases such TACI (CD267) and BCMA (CD269)) expressed predominantly as Sjoegren syndrome, Rheumatoid arthritis (RA), Multiple on B cells, although activated T cells also express BAFF-R. sclerosis (MS) and Systemic Lupus Erythematosus (SLE). BAFF BAFF is a master regulator of peripheral B cell survival, and has also increased levels in some lymphoid cancers.

Recombinant Proteins 1 Processed human BAFF can either remain as a trimer, which is usual for TNF family ligands, or assemble into 60-mer composed of 20

trimers. Mouse BAFF 60-mer has been identified in the serum of BAFF nm 490 OD transgenic mice. Despite the predominant functional role of processed BAFF in vivo, membrane-bound BAFF might also play a role. Indeed, 0 0.0001 0.001 0.01 100 1000 10 0.1 soluble BAFF (3-mer) can trigger BAFF-R but not TACI or BCMA, whereas 1 oligomeric forms of BAFF (BAFF 60-mer), which mimic membrane- ng/ml bound BAFF, activate all BAFF receptors. FIGURE: BAFF, Soluble (human) (60-mer) (AG-40B-0112) binds and activates BCMA receptor.

PRODUCT NAME PID SIZE SOURCE ENDOTOXIN SPECIES BAFF (aa134-285), Soluble (human) (rec.) AG-40B-0016 10 μg | 3 x 10 μg E. coli <0.01EU/μg Hu, Ms BAFF, Soluble (human) (60-mer) (rec.) AG-40B-0112 10 μg | 3 x 10 μg E. coli <0.01EU/μg Hu, Ms (highly active) Fc (human):BAFF (human) (rec.) AG-40B-0120 10 μg | 3 x 10 μg | CHO cells <0.01EU/μg Hu, Ms 500 μg BAFF, Soluble (mouse) (rec.) AG-40B-0022 10 μg | 3 x 10 μg HEK 293 cells <0.01EU/μg Ms, Hu BAFF-R (human):Fc (human) (rec.) AG-40B-0027 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms BCMA (human):Fc (human) (rec.) AG-40B-0080 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms BCMA (mouse):Fc (human) (rec.) AG-40B-0076 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Ms, Hu TACI (human):Fc (human) (rec.) AG-40B-0079 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms

Antibodies BULK available

PRODUCT NAME PID SIZE ISOTYPE APPLICATION SPECIES BAFF (human), mAb (1-35-1) AG-20B-0037 100 μg Rt IgG2a FACS Hu BAFF (human), mAb (2.81) AG-20B-0018 100 μg Rt IgG2b ELISA, IP Hu BAFF (human), mAb (blocking) (4.62) AG-20B-0017 100 μg Rt IgG2a ELISA, FUNC (Blocking), IP Hu BAFF (human), mAb (blocking) (4.62) (Biotin) AG-20B-0017B 100 μg Rt IgG2a ELISA, FUNC (Blocking), IP Hu BAFF-R (human), mAb (HuBR9.1) AG-20B-0016 100 μg Ms IgG1 FACS Hu BAFF-R (human), mAb (HuBR9.1) (Biotin) AG-20B-0016B 100 μg Ms IgG1 FACS Hu

www.adipogen.com BAFF-R (mouse), mAb (9B9) AG-20B-0034 100 μg Rt IgG2b FACS, FUNC (Depletion) Ms BAFF-R (mouse), mAb (9B9) (PF) AG-20B-0034PF 100 μg Rt IgG2b FACS, FUNC (Depletion) Ms BCMA (human), mAb (ANC3B1) ANC-269-020 100 μg Ms IgG1 ELISA, FACS Hu TACI (mouse), mAb (1A-10) AG-20B-0035 100 μg Rt IgG2a FACS Ms TACI (mouse), mAb (1A-10) (Biotin) AG-20B-0035B 100 μg Rt IgG2a FACS Ms

APPLICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; FORMULATION: PF = Preservative free 8 IHC: Immunohistochemistry IP: Immunoprecipitation; WB: Western blot SPECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; Prm = Primate BAFF ELISA Kit and Detection Set www.adipogen.com

BAFF, Soluble (human) BAFF, Soluble (human) ELISA Kit (hypersensitive) Matched Pair Detection Set AG-45B-0001-KI01 96 wells AG-46B-0001 1 Set SPECIES REACTIVITY: Human SPECIES REACTIVITY: Human SENSITIVITY: 8 pg/ml SENSITIVITY: 16 pg/ml RANGE: 15.6 to 1000 pg/ml RANGE: 16 to 1000 pg/ml DETECTION TYPE: Colorimetric DETECTION TYPE: Colorimetric ASSAY TYPE: Sandwich ASSAY TYPE: Sandwich SAMPLE TYPE: Cell Culture Supernatant, SAMPLE TYPE: Cell Culture Supernatant, Plasma, Serum Plasma, Serum

LIGAND: RECEPTORS: APRIL [TNFSF13] TACI [TNFRSF13B] BCMA [TNFRSF17]

APRIL (CD256) is a cytokine that belongs to the TNF super- regulation of tumor cell growth and is involved in monocyte/ family and binds to TACI (CD267) and BCMA (CD269). It macrophage-mediated immunological processes. APRIL pro- also interacts with heparan sulfate proteoglycans that duced by neutrophils sustains local antibody production by induces APRIL oligomerization, which is a prerequisite for the creating plasma cell niches in mucosa-associated lymphoid triggering of its effects on BCMA or TACI. It is implicated in the tissue (MALT).

Blocking Recombinant Antibody APRIL (mouse), mAb (rec.) (blocking) (Apry-1-1) AG-27B-0001-C100 100 μg AG-27B-0001PF-C100 Preservative Free 100 μg AG-27B-0001B-C100 Biotin 100 μg

CLONE: Apry-1-1. ISOTYPE: Mouse IgG2b. IMMUNOGEN: Mouse APRIL (aa 98-232). SPECIFICITY: Recognizes mouse APRIL. APPLICATION: ELISA, IP, FUNC (Inhibits binding of mouse APRIL to mouse BCMA and TACI). FIGURE: Binding of mouse APRIL to mouse BCMA is inhibited by anti-APRIL (mouse), mAb (rec.) (blocking) (Apry-1-1) (Prod. No. AG-27B-0001).

Oligomeric MegaAPRIL Proteins

PRODUCT NAME PID SIZE SOURCE ENDOTOXIN SPECIES MegaAPRIL, Soluble (human) (H98) (rec.) AG-40B-0088 10 μg | 3 x 10 μg HEK 293 cells <0.01EU/μg Hu, Ms MegaAPRIL, Soluble (human) (rec.) AG-40B-0017 10 μg | 3 x 10 μg HEK 293 cells <0.01EU/μg Hu, Ms MegaAPRIL, Soluble (mouse) (H98) (rec.) AG-40B-0035 10 μg | 3 x 10 μg HEK 293 cells <0.01EU/μg Ms, Hu MegaAPRIL, Soluble (mouse) (rec.) AG-40B-0089 10 μg | 3 x 10 μg HEK 293 cells <0.02EU/μg Ms, Hu

For updated prices and additional information visit www.adipogen.com or contact your local distributor. 9 LIGAND: RECEPTORS: RANKL [TNFSF11] RANK [TNFRSF11] OPG [TNFRSF11B]

RANKL (TRANCE; CD254) is an osteoclast differentiation and regulator of interactions between T cells and dendritic cells activation factor and is involved in osteoclastogenesis, binding and is involved in the regulation of the T cell-dependent to RANK (CD265). It augments the ability of dendritic cells immune response. RANKL plays an important role in the to stimulate naive T cell proliferation. It is an important progression of breast cancer.

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES RANK (human):Fc (human) (rec.) AG-40B-0018 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms RANK (mouse):Fc (human) (rec.) AG-40B-0092 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms Fc (human):RANKL, Soluble (mouse) (rec.) AG-40B-0059 10 μg HEK 293 cells <0.01EU/μg Ms ANTIBODIES PID SIZE SOURCE APPLICATION SPECIES RANK (ectodomain) (human), pAb AG-25A-0021 50 μg Rb FACS, WB Hu RANKL (human), pAb AG-25A-0016 100 μg Rb ELISA, FACS, WB Hu

LIGAND: RECEPTOR: GITRL [TNFSF18] GITR [TNFRSF18]

GITRL (Glucocorticoid-induced TNF receptor ligand; AITRL; TL6) GITRL is implicated in development of autoimmune diseases and is preferentially expressed by regulatory T cells. It is constitutively in the immune response against infectious pathogens and tumors. expressed and released as soluble form by solid tumors and Binding to GITR (AITR; CD357) is important in regulating various hematopoietic malignancies. GITRL causes differentia- T cell proliferation and TCR-mediated apoptosis. GITR serves tion of osteoclasts, activation of macrophages, but also alteration as a negative regulator for NK cell activation. It mediates of carcinoma and leukemia cells and influences apoptosis. NF-B activation via the TRAF2/NIK pathway.

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES GITR (human):Fc (human) (rec.) AG-40B-0028 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu GITR (mouse):Fc (human) (rec.) AG-40B-0002 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Ms GITRL, Soluble (human) (rec.) AG-40A-0019 50 μg HEK 293 cells <0.1EU/μg Hu GITRL, Soluble (human) (rec.) (His) AG-40A-0024 50 μg E. coli <1EU/μg Hu GITRL, Soluble (mouse) (rec.) AG-40A-0008 50 μg HEK 293 cells <0.1EU/μg Ms GITRL, Soluble (mouse) (rec.) (His) AG-40A-0009 50 μg E. coli <1EU/μg Ms ANTIBODIES PID SIZE SOURCE/ISOTYPE APPLICATION SPECIES GITR (human), mAb (AIT 158D) AG-20A-0017 50 μg | 100 μg Rt IgG2a ELISA, FACS Hu GITR (human), pAb AG-25A-0017 100 μg Rt ELISA, FACS Hu GITR (mouse), mAb (MGIT 02) AG-20A-0007 50 μg | 100 μg Rt IgG2a ELISA, FACS Ms GITR (mouse), mAb (MGIT 07) AG-20A-0008 50 μg | 100 μg Rt IgG2a ELISA, FACS, WB Ms GITR (mouse), pAb AG-25A-0001 100 μg Rt ELISA, FACS, WB Ms GITR (mouse), pAb (Biotin) AG-25A-0001B 50 μg Rt ELISA, FACS, WB Ms GITRL (human), pAb AG-25A-0023 100 μg Rb ELISA, WB Hu GITRL (mouse), mAb (MGTL 10) AG-20A-0009 50 μg | 100 μg Rt IgG2a ELISA, FACS Ms

www.adipogen.com GITRL (mouse), mAb (MGTL 15) AG-20A-0010 50 μg | 100 μg Rt IgG2a ELISA, FACS, WB Ms GITRL (mouse), pAb AG-25A-0002 100 μg Rb ELISA, FACS, WB Ms GITRL (mouse), pAb (Biotin) AG-25A-0002B 50 μg Rb ELISA, FACS, WB Ms GITRL (mouse), pAb AG-25A-0024 100 μg Rt ELISA, FACS, WB Ms

APPLICATIONS: FACS: Flow Cytometry; FUNC: Functional Application; ICC: Immunocytochemistry; FORMULATION: PF = Preservative free 10 IHC: Immunohistochemistry IP: Immunoprecipitation; WB: Western blot SPECIES: Hu = Human; Ms = Mouse; Rt = Rat; Rb = Rabbit; Prm = Primate LIGAND: RECEPTORS: TNF- [TNFSF2] TNF-R1 [TNFRSF1A] TNF-R2 [TNFRSF1B]

Tumor necrosis factor (TNF, cachexin or cachectin and formerly proteolytic activation which initiates the subsequent cascade known as tumor necrosis factor-) is a cytokine involved of caspases mediating apoptosis. TNF-R1 contributes to the in systemic inflammation and is a member of a group of induction of non-cytocidal TNF effects including anti-viral cytokines that stimulate the acute phase reaction. It binds to state and activation of the acid sphingomyelinase. TNF-R2 www.adipogen.com the receptors TNF-R1 (CD120a) and TNF-R2 (CD120b). TNF-R1 is is the high affinity receptor for TNF-. The TRAF1/TRAF2 the receptor for TNF- and LTS. The adapter molecule FADD complex recruits the apoptotic suppressors BIRC2 and BIRC3 recruits caspase-8 to the activated receptor. The resulting to TNFRSF1B/TNF-R2. This receptor mediates most of the death-inducing signaling complex (DISC) performs caspase-8 metabolic effects of TNF-.

PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES MegaTNF-, Soluble (human) (rec.) AG-40B-0019 10 μg HEK 293 cells <0.01EU/μg Hu, Ms TNF-, Soluble (human) (rec.) AG-40B-0006 10 μg | 50 μg | E. coli <0.01EU/μg Hu, Ms 3 x 50 μg TNF-R1 (human):Fc (human) (rec.) AG-40B-0074 50 μg | 3 x 50 μg HEK 293 cells <0.01EU/μg Hu, Ms ANTIBODY PID SIZE ISOTYPE APPLICATION SPECIES TNF- (human), mAb (J1D9) ANC-398-020 100 μg Ms IgG1 FACS, FUNC (Neu- Hu tralizing), WB

Other TNF Superfamily Members or Related Ligands

ANTIBODIES PID SIZE ISOTYPE APPLICATION SPECIES TNF NR CD27 (human), mAb (M-T271) ANC-176-020 100 μg Ms IgG1 ELISA, FACS Hu TNFRSF7 CD30 (human), mAb (AC10) ANC-179-020 100 μg Ms IgG2b FACS Hu TNFRSF8 CD70 (human), mAb (BU69) ANC-222-020 100 μg Ms IgG1 ELISA, FACS, FUNC Hu, TNFSF7 (Inhibition), ICC, IHC Prm NGFR (human), mAb (ANC271/3D7) ANC-271-020 100 μg Ms IgG1 ELISA, FACS Hu TNFRSF16 DR3 (human), mAb (ANC2D12) ANC-250-020 100 μg Ms IgG1 ELISA Hu TNFRSF25 PROTEINS PID SIZE SOURCE ENDOTOXIN SPECIES TNF NR CD27 (human) (rec.) (His) CHI-HR-200CD27 50 μg E. coli <0.1EU/μg Hu TNFRSF7 CD27 (human):Fc (human) (rec.) CHI-HF-210CD27 100 μg CHO cells <0.06EU/μg Hu TNFRSF7 CD70 (human)-muCD8 Fusion Protein ANC-537-020 25 μg CHO cells N/A Hu TNFSF7 NGFR (human)-muIg Fusion Protein ANC-527-020 25 μg CHO cells N/A Hu TNFRSF16 DR3 (human)-muIg Fusion Protein ANC-528-020 25 μg CHO cells N/A Hu TNFRSF25 DR6 (human):Fc (human) (rec.) AG-40B-0011 50 μg HEK 293 cells <0.01EU/μg Hu TNFRSF21 3 x 50 μg DR6 (mouse):Fc (human) (rec.) AG-40B-0062 50 μg HEK 293 cells <0.01EU/μg Ms TNFRSF21 3 x 50 μg EDA-A1, Soluble (human) (rec.) AG-40B-0106 10 μg E. coli <0.01EU/μg Hu, Ms N/A 3 x 10 μg EDAR (human):Fc (human) (rec.) AG-40B-0116 50 μg CHO cells <0.01EU/μg Hu, Ms N/A 3 x 50 μg Fn14 (human):Fc (human) (rec.) AG-40B-0034 50 μg HEK 293 cells <0.01EU/μg Hu, Ms TNFRSF12A 3 x 50 μg

TNF Ligands Enhancer – Facilitates the Oligomerization

PRODUCT NAME PID SIZE ENDOTOXIN SPECIES TNF Ligands Enhancer AG-35B-0001 50 μg <0.01EU/μg All

For updated prices and additional information visit www.adipogen.com or contact your local distributor. 11 (FLIP www.adipogen.com CATION: IP, WB. an epitope (aa 1-200) present in both short (FLIP an epitope (aa1-200)present inbothshort THE STANDARD G2B00-10 100μg 100μg 50μg AG-20B-0005-C100 FLIP, mAb(Dave-2) AG-20B-0056-C100 AG-20B-0056-C050 FLIP (human),mAb(NF6) (FLIP CLONE: CLONE: induced in inflammatory skin conditionsandbindto anovel tumornecrosis skin induced ininflammatory (2011) receptorfactor familymember, IGFLR1:A.A.Lobito, etal.; J. Biol. Chem. on functions regulatory exert T cell responses. maythat mIGFLandIGFL1produced potentially inthe skin expressed mostabundantlyonmouse T cells, suggesting is likelyanovel memberofthe TNF-R family. IGFLR1is amino acidsequence ofIGFLR1indicated thatthisreceptor family receptor 1(formerly AnalysisTMEM-149). ofthe to anovel andhighaffinity ficity receptor namedIGF-like withspeci-mIGFL andhumanIGFL1IGFL3interact IGFL1 expression samples. isincreased inpsoriaticskin wounding. humanonly skin orafter In responses inskin upregulated duringinflammation inmice andfurther skin 4) andoneinmice (mIGFL).mIGFLisexpressed innormal proteins consisting offour proteins inhumans(IGFL1to genefamilyisa newfamilyof factor-like Insulin-growth IGFLR1 LIT: IGFLR1 (human):Fc(human)(rec.) IGFL3 (human)(rec.) IGFL (mouse)(rec.) IGFLR1 (human),pAb(IN101) RTISPDSZ OREEDTXNSPECIES ENDOTOXIN SOURCE SIZE PID PROTEINS NIOYPDSZ OREAPPLICATION SPECIES SOURCE SIZE PID ANTIBODY Murine insulingrowthMurine (IGFL)andhumanIGFL1proteins factor-like are L S ) splice variantsofhumanandmouseFLIP. ) andlong(FLIP Dave-2. Dave-2. NF6. ICC, IHC, WB. ISOTYPE: – ISOTYPE: A NovelMemberofthe TNFRSF Antibodies forDeathReceptors DownstreamSignaling Mouse IgG1. Mouse L ) splice variantsofhumanFLIP. Rat IgG2a. Rat SPECIFICITY: SPECIFICITY: For local distributors please visitourwebsite. [email protected] FAX +41-61-926-60-49 +41-61-926-60-40 TEL Adipogen International EUROPE/REST OF WORLD Recognizes short G2B02 0 gR AS BHu FACS, WB Hu Hu Ms <0.05EU/μg Rb <0.5EU/μg 100μg HEK293cells <0.1EU/μg HEK293cells AG-25B-0026 50μg HEK293cells 10μg AG-40B-0087 10μg AG-40B-0090 AG-40B-0091 Recognizes S APPLICATION: ) andlong 286, APPLI- 18969 18969

G2B05-10 100μg 50μg AG-20B-0057-C100 AG-20B-0057-C050 Caspase-8 (human),mAb(C15) p18 subunit of human caspase-8. p18 subunitofhumancaspase-8. CLONE: For specificliterature references product and thendetected usingananti-FLAG® antibody (HRP). at 8μg/mlwithatwofold serialdilution)duringonehour(starting control, (human)(Prod. Omentin No. AG-40B-0042), were added ml overnight at room temperature. IGFL3(human)oranegative to itsligandIGFL3(human)(Prod. No. AG-40B-0090). METHOD: FIGURE: 0,5 1,5 2,5 3,5 [email protected] +1-858-457-8484 FAX +1-858-457-8383 TEL Adipogen Corp. NORTH & SOUTH AMERICA 0 1 2 3 C15. IGFLR1 (human):Fc (human)(Prod. No. AG-40B-0087) binds IGFLR1 (human):Fc was coated onanELISAplate at 1μg/ ISOTYPE: see www.adipogen.com Mouse IgG2b. Mouse APPLICATION: SPECIFICITY: Latest Insight [email protected] [email protected] +82-32-831-1470 FAX +82-32-858-1470 TEL Adipogen, Inc. SOUTH KOREA/ASIA Omenn Flag Omenn Flag IGFL3 h ICC, IP, WB. Recognizes the

JUL 2013 www.adipogen.com