NEW Products for Research

PROTEINS FOR EMERGING AVI-TAGGED BIOTINYLATED NEW SMALL MOLECULES IMMUNE CHECKPOINT TARGETS RECOMBINANT

APC 16673-34-0 NLRP3i BirA Enzyme Biotin O NH2 Signal 1 S MHC TCR-CD3 Biotin GLNDIFEAQKIEWHE O Complex O Cl N GLNDIFEAQKIEWHE H Signal 2 T Avi Tag O

ANTI-IDIOTYPE AND BIOSIMILAR RECOMBINANT MONOCLONAL CELL MARKERS INTERACTIVE ANTIBODIES ANTIBODIES RESOURCE TOOL

Cell Surface Markers = Intracellular Markers

NEW IMMUNOASSAYS SIMPLE PLEX™ ASSAYS LUMINEX® DISCOVERY FIXED PANELS

SINGLE-CELL WESTERNS RNASCOPE® ISH ASSAYS MEDIA & FBS

GMP CLOUDZ™ CELL SELECTION NEW LITERATURE NEW INTERACTIVE PATHWAYS & ACTIVATION KITS Immune Cell Isolation & Culture Quality Results Start at the Core Bio-Techne Immunoassays — Powered by R&D Systems CD4+ T Cell Subsets

Th2

Th1 Th9

Signal 1 B MHCII TCR-CD3 Complex Th17 Signal 2 Naïve CD4+ T Cell APC

Treg Th22

Tfh

Immunology T Y B O O K A C T I V I O M

L A S O Z L P U Z O R D S S W C R O E O I S R N D G F P I A

G

E U D O K S The Bio-Techne brands, R&D Systems®, Novus Biologicals®, Tocris®, ProteinSimple®, and ACD™, are always working hard to develop new products that will help to advance your research. Since we release new products weekly and the pace of research never slows down, we realize that it can be difficult to stay up-to-date on what new products or technologies are available. This guide is intended to highlight recently developed Bio-Techne products and resources that we feel are particularly relevant to immunology researchers.

Table of Contents

Bioactive Proteins for Emerging Immune Checkpoint Targets...... 2 Avi-Tagged Biotinylated Recombinant Proteins...... 6 New Tocris Small Molecules for Immunology Research...... 7 Cell Culture Media...... 8 Classic Fetal Bovine Serum...... 9 New R&D Systems and Novus Biologicals Antibodies for Immunology Research...... 10 Cell Markers Interactive Resource Tool...... 12 New Immunoassays for Immunology Research...... 14 Simple Plex™ Assays...... 15 New Luminex® Performance Discovery Panels...... 16 Single-Cell Westerns...... 17 RNAscope® ISH Assays for Inflammation, Immuno-Oncology, and Viral Pathogen Research...... 17 GMP-grade Immune Cell Culture Reagents...... 18

2 Bioactive Proteins for Emerging Immune Checkpoint Targets

Tumor cells frequently overexpress B7 family proteins that negatively regulate T cell activation, which allows them to evade detection by the host’s . As a result, researchers have focused on targeting these negative regulatory immune checkpoint pathways for cancer immunotherapy. Utilizing monoclonal antibodies directed against the T cell co-inhibitory receptors, CTLA-4, PD-1, or the PD-1 ligand, PD-L1, clinicians have seen remarkable success in some cancer patients. Unfortunately, they have also found that these antibodies are only effective in a minority of patients and some patients that initially show a positive response to treatment with these antibodies, can become resistant or relapse due to an upregulation of other immune checkpoint pathways. Therefore, additional immune checkpoint regulators that may serve as immunotherapeutic targets, either alone or in combination, are being sought. Bio-Techne is focused on developing new reagents for investigating potential next generation immune checkpoint targets. Our current portfolio includes R&D Systems bioactive recombinant proteins for studying members of the Butyrophilin, VSTM, VSIG, CEACAM, AMIGO, and SIRP families.

Butyrophilins

Butyrophilins are a novel class of co-stimulatory/co-inhibitory molecules that are structurally related to the B7 family and appear to have similar immunomodulatory functions. To date, 13 human butyrophilin proteins have been described, with most acting through unidentified receptors to inhibit T cell proliferation and production. The parallels between the butyrophilin proteins and the B7 family suggest that these proteins may also serve as drug targets for immune checkpoint blockade.

Butyrophilins Butyrophilins Molecule Species Source Tag Cat # Molecule Species Source Tag Cat # Human NS0 None 8467-BT BTNL2 Mouse NS0 None 8605-BT BTN1A1 Human NS0 Biotin BT8467 BTNL3 Human CHO Fc 9658-BT Mouse NS0 None 8540-BT Mouse NS0 Fc 9590-BT BTNL4 BTN2A1 Human HEK293 None 9058-BT Mouse NS0 His 9594-BT Human HEK293 Fc 8918-BT BTNL6 Mouse CHO None 9638-BT BTN2A2 Human HEK293 Fc, Biotin BT8918 Human HEK293 Fc 9359-BT BTNL8 Mouse NS0 None 8997-BT Human HEK293 Fc, Biotin BT9359 Human HEK293 Fc 8539-BT Human HEK293 Fc 9659-BT BTN3A1/CD277 BTNL9 Human HEK293 Fc, Biotin BT8539 Mouse NS0 Fc 9555-BT Human CHO None 9514-BT Human HEK293 Fc 10014-BT BTN3A2 BTNL10 Human CHO Biotin BT9514 Mouse NS0 Fc 1444-BT

BTN3A1 Inhibits Anti-CD3-induced IL-2 Production by A B Human T Cells in a Manner Similar to B7-H1/PD-L1. Human T cells were incubated with an immobilized Mouse Anti-Human CD3e Monoclonal Antibody (R&D Systems, Catalog # MAB100; 1 µg/mL) and the indicated ) R NR ) R NR concentrations of (A) Recombinant Human BTN3A1/ 40 40 R NR CD277 Fc Chimera (R&D Systems, Catalog # 8539-BT) or 190 (B) Recombinant Human B7-H1/PD-L1 Fc Chimera (R&D Systems, Catalog # 156-B7). IL-2 secretion was 92.5 measured in cell culture supernatants using the Human 66 ® 55 IL-2 Quantikine ELISA Kit (R&D Systems, Catalog # 43 D2050). The ED50 for this effect is typically 1-5 µg/mL for 36 Recombinant Human BTN3A1/CD277 and 2-10 µg/mL for 29

IL-2 Production (O.D. IL-2 Production (O.D. IL-2 Production Recombinant Human B7-H1/PD-L1. The purity of (A) 21 18.4 Recombinant Human BTN3A1/CD277 (1 µg/lane) and (B) 12.4 Recombinant Human B7-H1/PD-L1 (1 µg/lane) was 6.3 assessed by SDS-PAGE analysis under reducing (R) and non-reducing (NR) conditions and visualized by silver staining. Recombinant Human BTN3A1/Fc (g/mL) Recombinant Human B7-H1/Fc (g/mL)

Learn more | rndsystems.com/butyrophilins and rndsystems.com/butyrophilinwebinarshort 3 VSTM Family Proteins

There are 8 members of the V-set and transmembrane domain-containing (VSTM) protein family, which are structurally related to the B7 family proteins. R&D Systems scientists have found that many of these proteins have the R NR ability to suppress anti-CD3-induced cytokine secretion by human T cells, suggesting that they may also play a role in regulating T cell functions in vivo. VSTM Family Proteins Molecule Species Source Tag Catalog # Production (Mean O.D.) γ Production Human HEK293 His 10037-VT VSTM2A IFN- Mouse NS0 His 2283-VT Human HEK293 Fc 10060-VT VSTM2B Human HEK293 His 10061-VT Recombinant Human VSTM4 (g/mL) Mouse NS0 Fc 10076-VT VSTM4 Inhibits Anti-CD3-induced IFN-g Production by Human CHO HA 2111-VT VSTM2L Human T Cells. Human peripheral blood mononuclear Mouse CHO HA 2152-VT cells were incubated with an immobilized Mouse Anti-Human CD3e Monoclonal Antibody (R&D Systems, Human CHO Fc 7898-TGB Catalog # MAB100) and the indicated concentrations of Human CHO His 9525-TG Recombinant Human VSTM4 Fc Chimera (R&D Systems, Catalog # 2086-VT). IFN-g secretion was measured in cell VSTM3/TIGIT/VSIG9 Human HEK293 Fc 9464-TG culture supernatants using the Human IFN-g Quantikine®

Cynomolgus Monkey HEK293 Fc 9380-TG ELISA Kit (R&D Systems, Catalog # DIF50). The ED50 for this effect is typically 1-10 µg/mL. The purity of Mouse NS0 Fc 7267-TG Recombinant Human VSTM4 (2 µg/lane) was assessed by VSTM4 Human CHO Fc 2086-VT SDS-PAGE analysis under reducing (R) and non-reducing (NR) conditions and visualized by silver staining.

VSIG Family Proteins

Several members of the V-set and immunoglobulin domain-containing (VSIG) protein family are now recognized as potential immuno-oncology targets. The most well-known member of this family, VSIG9/TIGIT/VSTM3, has been shown to have both T cell and natural killer cell inhibitory functions. Additionally, VSIG4 has been reported to negatively regulate T cell proliferation and IL-2 production and inhibit pro-inflammatory macrophage activation, while VSIG3 has been shown to inhibit human T cell functions through binding to the B7 family member, VISTA/PD-1H. Our own unpublished observations indicate that other VSIG proteins also negatively regulate T cell functions.

VSIG Family Proteins VSIG Family Proteins Molecule Species Source Tag Cat. # Molecule Species Source Tag Cat. # Human NS0 Fc 9708-VS VSIG4 Short Human CHO His 9816-VS VSIG1 Mouse NS0 Fc 9645-VS Isoform Human CHO Fc 9817-VS Human NS0 Fc 9815-VS Human CHO Fc 9200-VS VSIG2 Mouse NS0 Fc 9598-VS Fc, VSIG8 Human CHO BT9200 Biotin Human NS0 Fc 9229-VS VSIG3 Mouse NS0 Fc 9204-VS

Mouse NS0 Fc 9367-VS (RF) T Cell Proliferation Human CHO Fc 7898-TGB VSIG3 Human CHO Fc 10074-VS N-Terminus Human CHO His 9525-TG VSIG9/ VSIG3 Human HEK293 Fc 9464-TG Human CHO Fc 10075-VS TIGIT/ VSIG3 IgG -Fc (g/mL) C-Terminus 1 VSTM3 Cynomolgus HEK293 Fc 9380-TG VSIG3 Inhibits Anti-CD3-induced Human CD3+ T Cell Human NS0 Fc 4646-VS Monkey Proliferation. Human T cells were incubated with an Fc, Mouse NS0 Fc 7267-TG immobilized Mouse Anti-Human CD3e Monoclonal Antibody Human NS0 BT4646 Biotin (R&D Systems, Catalog # MAB100; 1 µg/mL) and the VSIG10 Mouse NS0 Fc 9604-VS VSIG4 indicated concentrations of Recombinant Human VSIG3 Fc Human NS0 His 9678-VS Chimera (R&D Systems, Catalog # 9229-VS). Cell Mouse NS0 Fc 4674-VS proliferation was assessed by a fluorometric assay using the redox-sensitive dye, Resazurin (R&D Systems, Catalog Mouse NS0 His 9747-VS # AR002). IgG-Fc controls did not alter anti-CD3-induced CD3+ T cell proliferation (data not shown). Learn more | rndsystems.com/VSIG8poster and rndsystems.com/VSIG3-VISTAposter 4 CEACAM Proteins

Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) are part of the CEA protein family consisting of CEACAMs and the pregnancy-specific glycoproteins (PSGs). These proteins are part of the immunoglobulin superfamily and have a wide variety of different functions in cell adhesion, , inflammation, , and cancer. They have been shown to bind to each other and have been described as receptors for TIM-3, galectins, and integrins. Additionally, CEACAM-1, CEACAM-5, and CEACAM-6 are now considered to be cancer biomarkers and potential targets for cancer immunotherapy.

CEACAM Proteins Molecule Species Source Tag Catalog #

CEACAM-1/CD66a Human NS0 His 2244-CM Human NS0 His 9868-CM CEACAM-3/CD66d Human CHO Fc 9985-CM CEACAM-5/CD66e Human NS0 His 4128-CM CEACAM-6/CD66c Human NS0 His 3934-CM CEACAM-7 Human CHO His 9010-CM Galectin-3 Binding (Mean O.D.) CEACAM-8/CD66b Human NS0 His 9639-CM

CEACAM-16 Human CHO HA 9885-CM CEACAM-18 Human HEK293 Fc 9869-CM Recombinant Human CEACAM-21 (g/mL) CEACAM-19 Human HEK293 Fc 9397-CM CEACAM-21 Binds to Galectin-3. Recombinant Human CEACAM-20 Human CHO Fc 9427-CM Galectin-3 (R&D Systems, Catalog #8259-GA) was coated at 1 µg/mL, 100 µL/well and the indicated concentrations CEACAM-21 Human CHO Fc 9870-CM of Recombinant Human CEACAM-21 Fc Chimera (R&D Sys- tems, Catalog # 9870-CM) were added. The concentration of Recombinant Human CEACAM-21 Fc Chimera that produces 50% of the optimal binding response is 0.07–0.7 µg/mL. AMIGO Proteins

AMIGO family proteins (AMIGO1-3) are cell adhesion molecules that exhibit homophilic and heterophilic binding properties. Relatively little is known about their functions, but our in-house data shows that AMIGO2 and AMIGO3 inhibit anti-CD3-induced IL-2 secretion by CD3+ T cells, suggesting that they may also regulate T cell functions in vivo.

AMIGO Family Proteins Molecule Species Source Tag Catalog # Human NS0 His 2565-AM AMIGO Mouse NS0 His 10084-AM Human CHO Fc 2080-AM AMIGO2 Mouse NS0 Fc 9424-AM Human CHO Fc 9508-AM AMIGO3 Mouse NS0 Fc 9425-AM IL-2 Secretion (Mean O.D.) IL-2 Secretion

Recombinant Human AMIGO3 (g/mL) AMIGO3 Inhibits Anti-CD3-induced IL-2 Production by Mouse T Cells. Mouse T cells were treated with the indicated concentrations of Recombinant Human AMIGO3 Fc Chimera (R&D Systems, Catalog # 9508-AM) in the presence of anti-CD3 antibody and IL-2 secretion was measured using the Mouse IL-2 Quantikine® ELISA Kit (R&D Systems, Catalog # M2000).

Learn more | rndsystems.com/AMIGOs 5 SIRP Proteins and the CD47 Receptor

The SIRPa/CD47 signaling pathway is a myeloid cell-specific phagocytosis checkpoint that has recently gained more interest as a potential therapeutic target. Researchers have shown that CD47 is overexpressed on tumor cells, allowing them to evade the host’s immune system by inhibiting their phagocytosis. Reagents blocking CD47 signaling have been shown to activate macrophages or dendritic cells and stimulate phagocytosis, leading to tumor cell elimination. Preclinical studies have also suggested that SIRPa or CD47 blockade, used in combination with other immune checkpoint inhibitors that target the adaptive immune system, may have synergistic anti-tumor effects.

SIRP Proteins & CD47 Molecule Species Source Tag Catalog # Human NS0 Fc 4670-CD CD47 Human NS0 His 9396-CD

Mouse NS0 Fc 1866-CD Human CHO Fc 4546-SA Human CHO Fc, Biotin BT4546 Human CHO His 9378-SA SIRPa/CD172a Blood Cell Adhesion Red Cynomolgus Monkey NS0 His 9519-SA

Mouse NS0 Fc 7154-SA Rat NS0 His 7307-SA Recombinant Protein (g/mL) SIRPb2 Human HEK293 Fc 9998-SB Recombinant Mouse SIRPa Activity is Suppressed by a Soluble Version of the CD47 Receptor. Red blood cells Human CHO Fc 4486-SBB SIRPg/CD172g from mouse whole blood were incubated with the Human HEK293 His 9999-SB indicated concentrations of immobilized Recombinant Mouse SIRPa Fc Chimera (R&D Systems, Catalog # 7154-SA) for 1 hour at 37 ˚C. The plate was washed to remove non-adhered cells and red blood cell adhesion was assessed (green line). When Recombinant Mouse SIRPa Fc Chimera was immobilized at 2 µg/mL, the effect could be antagonized by the addition of soluble Recombinant Mouse CD47 Fc Chimera (R&D Systems, Catalog # 1866-CD; blue line). Avi-Tagged Biotinylated Recombinant Proteins

Avi-Tag Biotinylation Biotinylated proteins can be powerful tools for assessing protein-protein on nym II interactions and screening therapeutic antibodies or inhibitors of these on interactions. Bio-Techne is currently focused on expanding our selection of II Avi-tagged biotinylated recombinant proteins. Avi-tagged biotinylated g recombinant proteins are produced by overexpressing the specific Avi-tagged protein in cells along with the BirA biotin ligase to generate recombinant protein that is specifically biotinylated on the lysine residue of the Avi-tag. This method of enzymatic protein biotinylation offers several advantages over chemical biotinylation, including: • Elimination of random protein biotinylation and heterogeneous products that may have compromised activity • High specificity, leading to the generation of a homogeneous product with high yield • Easier to control the extent of biotinylation lot-to-lot • Enhanced control of protein orientation in binding assays

Avi-Tagged Biotinylated Recombinant Proteins As our catalog of Avi-tagged Molecule Species Source Tag Catalog # biotinylated proteins is expanding B7-1/CD80 Human CHO Avi, Biotin, His AVI9050 weekly, please visit rndsystems.com/ CD155/PVR Human HEK293 Avi, Biotin, His AVI9174 Fcg RIIIA Human HEK293 Avi, Biotin, His AVI4325 products/active-biotinylated-proteins PD-1 Human HEK293 Avi, Biotin, His AVI8986 to view a complete, up-to-date product PD-L1 Human HEK293 Avi, Biotin, His AVI9049 listing. 6 New Tocris® Small Molecules for Immunology Research

Bio-Techne provides a wide range of industry-leading, high performance Tocris small molecules for use in immunology research, including those that target , the complement system, and cytokine receptors, immune cell signaling, and viruses. A list of the most recently developed small molecules for immunology research is shown below.

Inflammasomes Product Name Catalog # Description NLRP3 inflammasome inhibitor. Inhibits formation of the NLRP3 inflammasome and IL-1β release in LPS and 16673-34-0 NLRP3i 6774 ATP-stimulated mouse macrophages and cardiomyocytes in vitro. NLRP3 inhibitor. Binds to the ATP-binding motif of the NLRP3 NACHT domain and blocks NLRP3 inflammasome CY 09 6436 assembly and activation. Potent DPP IV inhibitor. Activates the NLRP1b inflammasome and selectively triggers pyroptosis in monocytes and Talabostat mesylate 3719 macrophages. Orally bioavailable INF 39 6341 Irreversible NLRP3 inhibitor. Inhibits IL-1β release, caspase activation, and pyroptosis in THP-1 cells. STING (Stimulator of ) Murine-selective STING agonist. Triggers a type I IFN response in murine macrophages, but does not activate human CMA 6677 STING signaling. STING antagonist. Covalently binds STING at Cys91, blocking palmitoylation and preventing assembly of STING H 151 6675 clusters. Decreases production in human and mouse cellsin vitro. Immune checkpoints (D)-PPA 1 6515 PD-1/PD-L1 interaction inhibitor. Binds to PD-L1. Inhibits tumor growth and prolongs survival time of mice in vivo. TLRs Potent TLR8 inhibitor . Stabilizes the TLR8 dimer in its resting state. Suppresses TLR8-mediated proinflammatory CU CPT 9a 6479 signaling in various cell lines and human primary cells. Toll-like receptor 4 (TLR4) signaling inhibitor. Binds to the intracellular domain of TLR4 and inhibits LPS-induced TAK 242 6587 cytokine production in vitro and in vivo.

o log o log o log

N O O O NH2 S O O Cl N H N O OH

O OH

For a complete listing of small molecules for immunology research | tocris.com/research-area/immunology-research 7 Cell Culture Media

Bio-Techne now offers Atlanta Biologicals™ cell culture media including both RPMI 1640 and DMEM, as well as cell culture serum, media supplements, and cell culture antibiotics. With over 40 years of research and development experience invested, Atlanta Biologicals cell culture portfolio provides both an unparalleled commitment to quality and exceptional product value.

Cell Culture Media Catalog # Cell Culture Media Catalog # RPMI 1640, 25mM HEPES, no glutamine M30050 DMEM, low glucose, GlutaminePlus, no sodium pyruvate M17650 RPMI 1640, L-glutamine M30150 DMEM, low glucose, GlutaminePlus, no sodium pyruvate, no phenol red M17750 RPMI 1640, 25mM HEPES, L-glutamine M30250 DMEM, high glucose, 25mM HEPES, no glutamine, sodium pyruvate M18250 RPMI 1640, L-glutamine, no phenol red M30350 DMEM, high glucose, 25mM HEPES, no glutamine, sodium pyruvate, M18350 no phenol red RPMI 1640, no glutamine M30450 DMEM, high glucose, no glutamine, sodium pyruvate, no phenol red M18450 RPMI 1640, 25mM HEPES, no glutamine, no phenol red M30550 DMEM, high glucose, 25mM HEPES, L-glutamine, sodium pyruvate, RPMI 1640, no glutamine, no phenol red M30650 M18550 no phenol red RPMI 1640, 25mM HEPES, L-glutamine, no phenol red M30750 DMEM, high glucose, L-glutamine, sodium pyruvate, no phenol red M18650 RPMI 1640, GlutaminePlus M30850 DMEM, high glucose, 25mM HEPES, GlutaminePlus, sodium pyruvate M18750 RPMI 1640, 25mM HEPES, GlutaminePlus M31050 DMEM, high glucose, 25mM HEPES, GlutaminePlus, sodium pyruvate, M18850 RPMI 1640, 25mM HEPES, GlutaminePlus, no phenol red M31350 no phenol red RPMI 1640, GlutaminePlus, no phenol red M31550 DMEM, high glucose, GlutaminePlus, sodium pyruvate M18950 DMEM, low glucose, no glutamine, sodium pyruvate M12150 DMEM, high glucose, GlutaminePlus, sodium pyruvate, no phenol red M19050 DMEM, low glucose, no glutamine, no sodium pyruvate, no phenol red M12250 DMEM, high glucose, 25mM HEPES, no glutamine, no sodium pyruvate M19550 DMEM, low glucose, 25mM HEPES, no glutamine, sodium pyruvate M12350 DMEM, high glucose, 25mM HEPES, no glutamine, no sodium pyruvate, M19650 no phenol red DMEM, high glucose, no glutamine, sodium pyruvate M12450 DMEM, high glucose, 25mM HEPES, L-glutamine, no sodium pyruvate, DMEM, low glucose, 25mM HEPES, no glutamine, sodium pyruvate, M19750 M12550 no phenol red no phenol red DMEM, high glucose, 25mM HEPES, GlutaminePlus, no sodium M19850 DMEM, low glucose, no glutamine, sodium pyruvate, no phenol red M12650 pyruvate DMEM, low glucose 25mM HEPES, glutamine, sodium pyruvate, DMEM, high glucose, 25mM HEPES, GlutaminePlus, no sodium M12750 M19950 no phenol red pyruvate, no phenol red DMEM, low glucose, glutamine, sodium pyruvate, no phenol red M12850 DMEM, low glucose, glutamine, sodium pyruvate M22150 DMEM, high glucose, L-glutamine, no sodium pyruvate, no phenol red M12950 DMEM, high glucose, L-glutamine, no sodium pyruvate M22250 DMEM, low glucose, 25mM HEPES, GlutaminePlus, sodium pyruvate M13050 DMEM, low glucose, 25mM HEPES, glutamine, sodium pyruvate M22350 DMEM, low glucose, 25mM HEPES, GlutaminePlus, sodium pyruvate, M13150 DMEM, high glucose, L-glutamine, sodium pyruvate M22450 no phenol red DMEM, high glucose, 25mM HEPES, L-glutamine, no sodium pyruvate M22550 DMEM, low glucose, GlutaminePlus, sodium pyruvate M13250 DMEM, high glucose, 25mM HEPES, L-glutamine, sodium pyruvate M22650 DMEM, low glucose, GlutaminePlus, sodium pyruvate, no phenol red M13350 DMEM, high glucose, no glutamine, no sodium pyruvate M22850 DMEM, low glucose, 25mM HEPES, no glutamine, no sodium pyruvate M13850 DMEM, high glucose, no glutamine, no sodium pyruvate, no phenol red M22950 DMEM, low glucose, 25mM HEPES, no glutamine, no sodium pyruvate, M13950 no phenol red DMEM, high glucose, GlutaminePlus, no sodium pyruvate, M32350 no phenol red DMEM, low glucose, no glutamine, no sodium pyruvate M14050 DMEM, high glucose, GlutaminePlus, no sodium pyruvate M32650 DMEM, low glucose, 25mM HEPES, glutamine, no sodium pyruvate M17050 DMEM, low glucose, 25mM HEPES, glutamine, no sodium pyruvate, M17150 no phenol red DMEM, low glucose, glutamine, no sodium pyruvate M17250 DMEM, low glucose, glutamine, no sodium pyruvate, no phenol red M17350 DMEM, low glucose, 25mM HEPES, GlutaminePlus, no sodium pyruvate M17450 DMEM, low glucose, 25mM HEPES, GlutaminePlus, no sodium pyruvate, M17550 no phenol red

Learn more | atlanta-biologicals.com

8 Classic Fetal Bovine Serum

Fetal bovine serum (FBS) is the ideal growth supplement for cell culture media and is generally preferred over other types of sera due to its high levels of nutrients and optimal combination of growth factors. Atlanta Biologicals FBS is produced utilizing strict manufacturing processes and quality assurance procedures, which guarantees unsurpassed quality and consistency from lot-to-lot.

Fetal Bovine Serum Product Size Regular (Catalog #) Heat-inactivated (Catalog #) 50 mL S12495 S12495H Fetal Bovine Serum - Optima 100 mL S12410 S12410H 500 mL S12450 S12450H 50 mL S11195 S11195H Fetal Bovine Serum - Premium 100 mL S11110 S11110H 500 mL S11150 S11150H 50 mL S11595 S11595H Fetal Bovine Serum - Premium Select 100 mL S11510 S11510H 500 mL S11550 S11550H

Other Bovine Serum Product Size Regular (Catalog #) Heat-inactivated (Catalog #) 50 mL S11295 S11295H Newborn Calf Serum 100 mL S11210 S11210H 500 mL S11250 S11250H 50 mL S11495 S11495H Calf Serum 100 mL S11410 S11410H 500 mL S11450 S11450H 50 mL S11995 S11995H Iron-Supplemented Calf Serum 100 mL S11910 S11910H 500 mL S11950 S11950H 50 mL S11395 S11395H Donor Bovine Serum 100 mL S11310 S11310H 500 mL S11350 S11350H

Reagents and Supplements In addition to media and serum, Bio-Techne also now offers Atlanta Biologicals media supplements, balanced salt solutions, biological buffers, cell freezing media, and more.

Learn more | atlanta-biologicals.com

9 New R&D Systems® and Novus Biologicals® Antibodies For Immunology Research

Bio-Techne offers an unparalleled selection of unlabeled and fluorochrome-conjugated R&D Systems and Novus Biologicals antibodies that are qualified for one or more of the following applications: blocking/ neutralization, ELISA, flow cytometry, immunocytochemistry/, , , Simple Western, and . Hundreds of world-renowned unique clones are available from the R&D Systems brand, many of which have been used to establish CD nomenclature through HLDA workshops. In addition, the Novus Biologicals brand includes an expansive collection of some of the most highly referenced antibody clones on the market. Most of these are conjugated to multiple different fluorochromes to provide a full range of options for multicolor experiments. A select list of new antibodies available from the R&D Systems and Novus Biologicals brands are shown in the sections that follow.

R&D Systems® Anti-Idiotype and Biosimilar Antibodies

Bio-Techne now offers anti-idiotype antibodies designed to bind specifically to the variable region of a monoclonal antibody drug, along with biosimilar antibodies that are intended to serve as research use only controls for our anti-idiotype antibodies. Biosimilar antibodies are expressed recombinantly using the same sequence as the original therapeutic antibody, and each anti-idiotype antibody is specifically designed to detect the biosimilar antibody it is targeting. The specificity and functionality of our anti-idiotype antibodies are confirmed using extensive in-house testing including bridging assays, flow cytometry, binding inhibition, and direct ELISA testing.

Anti-ID Anti-ID Anti-ID Drug Target

Non-inhibitory Antibody Drug Detects the drug-targeted complex Inhibitory/Neutraliing Detects total drug Antibody Drug Non-inhibitory Detects free drug (free and bound)

New Anti-Idiotype and Biosimilar Antibodies Molecule Species Clone Catalog # (Applications) Fluorochrome-conjugated Antibodies for Flow Cytometry (Cat. #s) Multi-species 972557 MAB9546 (E, WB) Adalimumab Multi-species 2235F MAB9616 (E, WB) Brentuximab Multi-species 2262C MAB9584 (E, FC, WB) FAB9575B, FAB9575G, FAB9575N, FAB9575R, FAB9575S, CD20 (Rituximab Biosimilar) Human Hu2 MAB9575 (CyTOF-ready, FC) FAB9575T, FAB9575U, FAB9575V FAB9576B, FAB9576G, FAB9576N, FAB9576R, FAB9576S, CD30/TNFRSF8 (Brentuximab Biosimilar) Human Hu8 MAB9576 (CyTOF-ready, FC) FAB9576T, FAB9576U, FAB9576V FAB9889B, FAB9889G, FAB9889N, FAB9889R, FAB9889S, CD52 (Alemtuzumab Biosimilar) Human Hu116 MAB9889 (CyTOF-ready, FC) FAB9889T, FAB9889U, FAB9889V Multi-species 2263B MAB9617 (E, WB) Gemtuzumab Multi-species 975155 MAB96171 (E, FC, WB) Ibritumomab Multi-species 974622 MAB9544 (E, WB) Rituximab Multi-species 2260A MAB9630 (BI, E, FC, WB) FAB9635B, FAB9635G, FAB9635N, FAB9635R, FAB9635S, Siglec-3/CD33 (Gemtuzumab Biosimilar) Human Hu9a MAB9635 (CyTOF-ready, FC) FAB9635T, FAB9635U, FAB9635V IC9677B, IC9677G, IC9677N, IC9677R, IC9677S, IC9677T, TNF-a (Adalimumab Biosimilar) Human Hu7 MAB9677 (B/N, FC) IC9677U, IC9677V

Application Key: BI Binding Inhibition B/N Blocking/Neutralization CyTOF-ready Mass cytometry E ELISA FC Flow Cytometry WB Western blot Learn more | rndsystems.com/products/anti-idiotype-antibodies and rndsystems.com/products/biosimilar-antibodies 10 R&D Systems® Recombinant Monoclonal Antibodies

Bio-Techne now offers recombinant monoclonal antibodies to minimize one potential source of experimental variability in your research. Our catalog includes a wide selection of recombinant antibodies from rabbit, mouse, and rat host species. The benefits of recombinant antibodies include: • Consistent and reproducible supply • Undergo the same validation testing as our traditional antibodies • Large quantities can be produced to meet your research needs

New Recombinant Monoclonal Antibodies New Recombinant Monoclonal Antibodies Molecule Species Clone Catalog # (Applications) Molecule Species Clone Catalog # (Applications) Angiopoietin-2 H 85816R MAB098R (E) MAB160R (B/N, CyTOF- H 49801R ready, FC, IHC) Angiopoietin-2 H 85834R MAB0984 (E) MAB1685R (CyTOF-ready, MAB154R (CyTOF-ready, CXCR3 M 220803R Axl H 108724R FC) Ecap, FC) MAB8109R (CyTOF-ready, MAB65764 (CyTOF-ready, R 868013R B7-H4 H 2318A FC) FC, IHC, WB)* MAB21651R (CyTOF-ready, MAB21541 (CyTOF-ready, CXCR4 M 247506R B7-H4 H/M 2319B FC, ICC, IHC) FC, IHC, WB)* MAB4227R (CyTOF-ready, BTN1A1/Butyrophilin H 2151B MAB84671 (FC, ICC/IF)* H 358426R FC, IHC) BTNL8 H 2187B MAB9359 (FC, SW, WB)* MAB42273R (CyTOF-ready, CXCR7 H 11G8R CCL2/JE/MCP-1 H 23007R MAB679R (B/N, Ecap, WB) FC, IHC) H 48607R MAB150R (CyTOF-ready, FC) MAB8399R (CyTOF-ready, R 896032R FC) MAB55381R (CyTOF-ready, M 475301R CCR2 FC) MAB83451R (CyTOF-ready, DC-SIGN/CD209 M MMD3R FC) MAB8368R (CyTOF-ready, R 890231R FC) EBOV GP V 993408 MAB9016 (WB) MAB1567R (CyTOF-ready, GDF-11/GDF-8 H 743835R MAB19582R (B/N, E) CCR4 H 205410R FC) GM-CSF H 3209R MAB215R (B/N, ICC, WB) MAB1802R (CyTOF-ready, CCR5 H CTC5R IFN-g H 25718R MAB285R (B/N, ICC) FC) IL-10 H 25209R MAB2171R (B/N, WB) MAB195R (CyTOF-ready, FC, H 53103R IHC) IL-12/IL-35 p35 H/M 27537R MAB1570R (WB) MAB590R (CyTOF-ready, FC, MAB227R (B/N, CyTOF- CCR6 M 140706R IL-6 Ra H 17506R IHC) ready, Ecap, FC, WB) MAB8320R (CyTOF-ready, LIGHT/TNFSF14 M 885310 MAB17942 (FC) R 876515R FC) MMP-9 H 4H3R MAB911R (IP, WB) MAB197R (B/N, CyTOF- CCR7 H 150503R MAB1059R (CyTOF-ready, ready, FC) H 131411R FC) MAB3478R (CyTOF-ready, NKG2A/CD159a H 314305R MAB68671 (CyTOF-ready, FC) M 2097C CCR10 FC) MAB2815R (CyTOF-ready, M 248918R MAB1562 (B/N, CyTOF- FC) PD-L1/B7-H1 H 2340D ready, FC, WB) H 50040R MAB3833R (Ecap) RORgt/RORC2/NR1F3 H/M 1181A MAB61093 (FC)* CD14 MAB3832R (B/N, H 134620R MAB72671 (CyTOF-ready, CyTOF-ready, FC) TIGIT M 2190A FC) CD19 R 771404R MAB7489R (IHC) MAB2365R (CyTOF-ready, H 344823R CD28 H 37407R MAB342R (A, WB) FC) TIM-3 MAB4625R (CyTOF-ready, MAB1529R (CyTOF-ready, CD37 H 424925R M 215008R FC) FC)* CD97 H 380905R MAB25291R (B/N, Ecap) MAB2929R (CyTOF-ready, TIM-4 H 921832R FC) C-Reactive Protein/CRP H 232007R MAB17071R (Ecap) TNF-a H 2342C MAB2103 (B/N, WB) CTLA-4 H 2188A MAB3252 (FC, ICC) MAB71261R (CyTOF-ready, MAB8110R (CyTOF-ready, VISTA/B7-H5/PD-1H H 730804R CXCR2/IL-8 RB R 866614R FC) FC)

*Several Alexa Fluor®-conjugated forms of this antibody are also available. Species Key: H Human M Mouse R Rat Application Key: A Agonist Activity B/N Blocking/Neutralization CyTOF-ready Mass Cytometry E ELISA Ecap ELISA Capture FC Flow Cytometry ICC Immunocytochemistry IP Immunoprecipitation WB Western Blot Learn more | rndsystems.com/products/recombinant-antibodies 11 Additional New R&D Systems® Antibodies for Immunology Research

Additional New R&D Systems Antibodies Additional New R&D Systems Antibodies Molecule Species Clone Catalog # (Applications) Molecule Species Clone Catalog # (Applications) CD2 H 299808 MAB18562 (A, WB) Indoleamine 2,3-dioxy- H 998743 MAB60301 (IHC) genase/IDO CD28 H 987701 MAB3421 (IHC) V 995047 MAB100061 (E) MAB5405 (CyTOF-ready, FC, Japanese Encephalitis CD72 H 982405 ICC) virus NS1 V 995026 MAB100062 (E) MAB20671 (CyTOF-ready, Ly-6G (Gr-1) M 1A8 MAB10371 (FC) CD90/Thy1 H 783922 FC) MAVS H 981240 MAB9694 (IHC) V 988110 MAB9779 (E) H 610615 MAB22291 (B/N) Chikungunya Virus E1 V 988133 MAB97791 (E) Nectin-2/CD112 H 610611 MAB22292 (B/N, ICC) V 988122 MAB97792 (WB) MAB10543 (B/N, OX40/TNFRSF4 H 977974 CTLA-4 H 922137 MAB3254 (B/N) CyTOF-ready, FC) V 973602 MAB9438 (E) P2X7/P2RX7 H 545230 MAB9790 (ICC) Dengue Virus 1 NS1 V 973604 MAB94381 (E) PD-L2/B7-DC H 988708 MAB12241 (FC) Dengue Virus 1/3 NS1 V 969977 MAB94402 (E) Perforin H 1001103 MAB8011 (IHC, WB) V 969973 MAB94401 (E, WB) TBK1 H Polyclonal AF9934 (IHC) Dengue Virus NS1 V 971305 MAB94441 (E, WB) TDAG8/GPR65 H 976128 MAB10077 (ICC, WB) V 971332 MAB94442 (E) MAB94182 (B/N, VSIG8 H 961829 CyTOF-ready, FC) MAB9494 (CyTOF-ready, FC, Fc mu R/FAIM3 H/M/R 992338 WB) VSIG10 H 988622 MAB9969 (ICC) MAB10082 (CyTOF-ready, Zika Virus (H/PF/2013) FFAR2/GPR43 H 1000624 V 990801 MAB10009 (E, WB) FC) Envelope H 999916 MAB10050 (E, SW, WB) Zika Virus (H/PF/2013) V 990844 MAB100091 (E, WB) Ig Kappa Light Chain Envelope H 999947 MAB100501 (E, SW, WB) Zika Virus (H/PF/2013) H 1000011 MAB10049 (E, SW, WB) V 990948 MAB9920 (E) Ig Lambda Light Chain NS1 H 1000040 MAB100491 (E, SW, WB) Zika Virus (H/PF/2013) V 990907 MAB99201 (E) IL-21 H Polyclonal AF15001 (WB) NS1

Species Key: H Human M Mouse V Viral Application Key: A Agonist Activity B/N Blocking/Neutralization CyTOF-ready Mass Cytometry E ELISA FC Flow Cytometry ICC Immunocytochemistry IHC Immunohistochemistry IP Immunoprecipitation SW Simple Western WB Western Blot For a complete listing of R&D Systems antibodies for immunology research | rndsystems.com/research-area/immunology

Cell Markers Interactive Resource Tool

Explore our new Cell Markers Interactive Resource Tool Natural Killer Cell Markers on the R&D Systems website to view the markers that are most commonly used in the scientific literature to identify and characterize different immune cell types or cell type-specific subsets. This tool will not only help you find appropriate cell markers, but it will also let you browse Bio-Techne’s selection of highly sensitive and specific antibodies that are available from the R&D Systems and Novus Biologicals brands. When applicable, the pages for different cell types show flow cytometry or IHC data generated using our antibodies.

Learn more | rndsystems.com/cellmarkers

12 New Novus Biologicals® Antibodies for Immunology Research

New Novus Biologicals Antibodies New Novus Biologicals Antibodies Molecule Species Clone Catalog # (Applications) Molecule Species Clone Catalog # (Applications) H/M/R A2-B10 NBP2-76901 (FC, IHC, WB) NBP2-67722 (FC, ICC/IF, CD81 H/M/R SN206-01 a1-Acid Glycoprotein IHC, WB) H/M/R A4-G2-A3 NBP2-76902 (FC, IHC, WB) CXCR5 H/M/R JB11-40 NBP2-75460 (FC, IHC, WB) NBP2-67417 (FC, ICC/IF, IHC, B7-2/CD86 H/CM/R SJ20-00 IP, WB) Chymase/CMA1 H/M/R JB74-32 NBP2-75441 (IHC, WB) NBP2-67456 (ICC/IF, IHC, IP, NBP2-76879 (ICC/IF, IHC, Bim H SU0318 HLA B H/M/R 11C2 WB) WB) NBP2-67700 (FC, IHC, IP, HLA Class I H/M Polyclonal NBP2-75928 (FC, IHC, WB) CCR2 H/M/R SN707 WB) Integrin a2/CD49b H CL7318 NBP2-76483 (IHC, WB) NBP2-67324 (ICC/IF, IHC, IP, CCR7 H/M/R SR36-04 Integrin b4/CD104 H/M/R CL6972 NBP2-76524 (IHC, WB) WB) NBP2-76843 (FC, ICC/IF, NBP2-75435 (FC, IHC, IP, IRAK2 H JG36-23 CD3g H/M JB38-29 IHC, WB) WB) NBP2-76906 (FC, ICC/IF, NBP2-75434 (FC, ICC/IF, Stathmin 1 H 110 CD27/TNFRSF7 H/R JB40-98 IHC, WB) IHC, WB) NBP2-75719 (FC, ICC/IF, NBP2-75436 (ICC/IF, IHC, ZAP70 H JU08-39 CD46 H/R JB25-49 IHC, IP, WB) WB) CD47 H CL6913 NBP2-76511 (IHC)

Species Key: H Human M Mouse CM Cynomolgus Monkey R Rat Application Key: FC Flow Cytometry ICC/IF Immunocytochemistry/Immunofluorescence IHC Immunohistochemistry IP Immunoprecipitation WB Western Blot

For a complete listing of Novus Biologicals antibodies for immunology research | novusbio.com/research-areas/immunology

13 New Immunoassays for Immunology Research

Bio-Techne’s portfolio includes single analyte Quantikine® ELISA Kits and DuoSet® ELISA Development Systems from the R&D Systems brand, quantitative multiplex Luminex® Assays from the R&D Systems brand, and fully automated, single or multianalyte Simple Plex™ assays run on Ella™ from the ProteinSimple brand. The following sections outline the new products that are available in each of these product categories.

Quantikine® ELISA Kits

• Complete, ready-to-use kits • Exhaustively tested for superior performance and reproducibility • The most referenced in the scientific literature

New Quantikine ELISA Kits Molecule Species Catalog # onug on noy u Axl Human DAXL00 Granzyme B Human DGZB00 IFN-b Human DIFNB0 IGF-II Mouse/Rat MG200 Progranulin Mouse MPGRN0 u

lu Serum EDTA Plasma o llo

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Heparin Plasma Cell Culture Supernate

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Assay Linearity is an Important Measure of Immunoassay Accuracy. Spiked serum, EDTA plasma, heparin plasma, and cell culture supernatant samples were serially diluted and assayed for human Axl using either R&D Systems Human Axl Quantikine ELISA Kit (R&D Systems, Catalog # DAXL00) or a competitor’s . All samples measured with the Human Axl Quantikine ELISA Kit had recovery values between 90–110% of the neat sample and displayed acceptable assay linearity (blue lines), while the percent recovery of diluted samples measured with the competitor’s kit had a range of 145–275% (orange lines), suggesting that interfering factors were preventing accurate measurement of the target analyte.

14 DuoSet® ELISA Development Systems

• Contain all the essential components required to develop an immunoassay for a specific target, but require the user to set up the assay • Economical and adaptable to multiple platforms

New DuoSet ELISA Development Systems New DuoSet ELISA Development Systems New DuoSet ELISA Development Systems Molecule Species Catalog # Molecule Species Catalog # Molecule Species Catalog # 5'-Nucleotidase/CD73 Human DY5795 DPPII/QPP/DPP7 Human DY3438 IFN-a2 Human DY9345 Arrestin Human DY9925 EBI3 Human DY6456 IL-21 Human DY8879 B7-H4 Human DY6576 ECM1 Human DY3937 Irisin/FNDC5 Human DY9420 BPI Human DY7468 EGF R Mouse DY1280 JE/MCP-1/CCL2 Rat DY3144 CD21 Human DY4909 Epiregulin Human DY1195 PDGF-AB Mouse/Rat DY8460 CD83 Human DY2044 FABP1/L-FABP Mouse/Rat DY1565 PSGL-1/CD162 Human DY3345 CD157 Human DY4736 Galectin-4 Human DY1227 Serpin B2 Human DY8550 Cochlin Human DY5249 Glypican 1 Human DY4519 SLPI Mouse DY1735 C-Peptide Human DY9625 Hepsin Human DY4776 UCH-L1/PGP9.5 Human DY6007 CTGF/CCN2 Human DY9190 HIV-1 Gag p24 Viral DY7360

Learn more | rndsystems.com/ELISAs

Simple Plex™ Assays

Simple Plex assays are fully automated, single or multianalyte assays run on Ella. Ella has three Simple Plex cartridges, either a 32x4 or a 16x4 cartridge for multiplexing, or a 72x1 cartridge for single analyte assays. In addition to the large selection of analytes that are currently available on the standard Simple Plex menu, we now also offer a 48-Digoxigenin (48-Dig) Simple Plex cartridge that gives researchers the freedom to choose the analytes that they want to detect in a Simple Plex assay. This new 48-Dig cartridge expands your assay options beyond the preset menu, offering researchers the opportunity to build their own sandwich assays on Ella. • Single and multianalyte assay formats are available • Unprecedented reproducibility • Wide dynamic range and sensitivity • Hands-free, fast time to results

Learn more | proteinsimple.com/ella

15 New Luminex® Performance Discovery Panels nly

To help researchers make their discoveries faster, Bio-Techne is now offering

Human Luminex Performance Discovery Assay fixed panels from the n on

R&D Systems brand. These panels are available off-the-shelf and are ready on noy g nly to ship upon order. As an alternative, researchers can select any combination u noy of the analytes that are part of the Human XL Cytokine Discovery Panel umn (R&D Systems, Catalog # FCSTM18) to make their own custom assay.

Magnetic Luminex® Performance Assay Fixed Panels Panel Analytes Catalog # • CCL2/MCP-1 • CCL3/MIP-1a • CCL4/MIP-1b • CCL5/RANTES • CCL11/Eotaxin • CXCL1/GROa Human Fixed Chemokine Discovery 8-plex LKTM012 • CXCL10/IP-10 • IL-8/CXCL8 • CCL2/MCP-1 • CCL3/MIP-1a • CCL4/MIP-1b • CCL5/RANTES • CCL11/Eotaxin • CCL19/MIP-3b • CCL20/MIP-3a • CD40 Ligand • Fractalkine/CX3CL1 • CXCL1/GROa • CXCL2/GROb • CXCL10/IP-10 • EGF Human Fixed Cytokine Discovery 45-plex • FGF basic • Flt-3 Ligand • G-CSF • GM-CSF • Granzyme B • IFN-a • IFN-b • IFN-g • IL-1a • IL-1b • IL-1ra LKTM014 • IL-2 • IL-3 • IL-4 • IL-5 • IL-6 • IL-7 • IL-8/CXCL8 • IL-10 • IL-12 p70 • IL-13 • IL-15 • IL-17/IL-17A • IL-17E/IL-25 • IL-33 • PD-L1/B7-H1 • PDGF-AA • PDGF-AB/BB • TGF-a • TNF-a • TRAIL • VEGF Human Fixed Cytokine Discovery 14-plex • IFN-a • IFN-g • IL-1a • IL-1b • IL-1ra • IL-2 • IL-3 • IL-4 • IL-6 • IL-7 • IL-10 • IL-15 • IL-33 • VEGF LKTM011 • CCL3/MIP-1a • CCL19/MIP-3b • CD40 Ligand • Fractalkine/CX3CL1 • CXCL2/GROb • EGF • FGF basic Human Fixed Discovery 21-plex • Flt-3 Ligand • G-CSF • Granzyme B • IFN-b • IL-3 • IL-8/CXCL8 • IL-17E/IL-25 • IL-33 • PD-L1/B7-H1 LKTM013 • PDGF-AA • PDGF-AB/BB • TGF-a • TRAIL • VEGF • CCL2/MCP-1 • CCL3/MIP-1a • CCL4/MIP-1b • CD40 Ligand • CXCL10/IP-10 • GM-CSF • Granzyme B Human Fixed Immunotherapy Discovery • IFN-a • IFN-g • IL-1a • IL-1b • IL-1ra • IL-2 • IL-4 • IL-6 • IL-8/CXCL8 • IL-10 • IL-12 p70 • IL-13 • IL-15 LKTM010 24-plex • IL-17/IL-17A • IL-33 • PD-L1/B7-H1 • TNF-a Human Fixed Th1/Th2 Discovery 11-plex • GM-CSF • IFN-g • IL-1b • IL-2 • IL-4 • IL-5 • IL-6 • IL-10 • IL-12 p70 • IL-13 • TNF-a LKTM008 Human Fixed Th9/Th17/Th22 Discovery • CD40 Ligand • GM-CSF • IFN-g • IL-1b • IL-2 • IL-4 • IL-5 • IL-6 • IL-10 • IL-12 p70 • IL-13 • IL-15 LKTM009 17-plex • IL-17/IL-17A • IL-17E/IL-25 • IL-33 • MIP-3a • TNF-a

A B nn nn onnon gm onnon gm

I I Iβ I I I Iγ I I α nly nly

Quantitation of in Cell Culture Supernates from Activated Th1 and Th2 Cells. CD4+ T cells were isolated from peripheral blood mononuclear cells (PBMCs) using the MagCellect Human CD4+ T Cell Isolation Kit (R&D Systems, Catalog # MAGH102). Cells were either untreated (NT), treated with Recombinant Human IL-2 (R&D Systems, Catalog # 202-IL), Recombinant Human IL-12 (R&D Systems, Catalog # 219-IL), and Mouse Anti-Human IL-4 Monoclonal Antibody (R&D Systems, Catalog # MAB304) and activated with immobilized Mouse Anti-Human CD3e Monoclonal Antibody (R&D Systems, Catalog # MAB100) and soluble Mouse Anti-Human CD28 Monoclonal Antibody (R&D Systems, Catalog # MAB342) to induce Th1 differentiation, or treated with Recombinant Human IL-2 (R&D Systems, Catalog # 202-IL) and activated with Phytohemagglutinin-L (PHA) to induce Th2 differentiation. All stimulated cells were then treated with PMA (Tocris, Catalog # 1201; 10 ng/mL) and Ionomycin calcium salt (Tocris, Catalog # 1704; 500 ng/mL) for 24 hours after activation. Cell culture supernates were analyzed using the Magnetic Luminex Performance Human Fixed Th1/Th2 Discovery 11-plex Panel (R&D Systems, Catalog # LKTM008). + = Values above the limits of the standard curve • = Stimulating cytokine

Learn more | rndsystems.com/luminexdiscoveryperformanceassays 16 Single-Cell Westerns

Milo™ is the world’s first single-cell Western platform. He measures protein expression in thousands of single cells in a single run, for up to 12 targets per cell. Milo can be utilized to help alleviate many of the common problems encountered with flow cytometry experiments including potentially limited target accessibility, poor antibody quality, and off-target antibody binding affecting reproducibility of the data. Milo performs single-cell protein analysis without the need to fix and permeabilize cells, making it easier to detect intracellular proteins. Additionally, Milo allows you to utilize antibodies selected from a large catalog of traditional Western blot antibodies, providing greater assay flexibility and enabling the detection of proteins that don’t have high-quality, flow cytometry-validated antibodies. He also incorporates a Learn more | size-based separation step prior to antibody probing to provide molecular proteinsimple.com/milo weight and antibody specificity information that will give you confidence in the accuracy of target detection.

RNAscope® ISH Assays for Inflammation, Immuno-Oncology, and Viral Pathogen Research

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g olgo o The RNAscope technology is a novel in situ hybridization method that is designed to detect target RNA in intact cells by using proprietary probes that simultaneously amplify target signal sequences while suppressing background noise from non-specific hybridization. This technology is particularly useful when your target is either a novel with no commercial antibody available, a secreted molecule that requires identifying the cellular source, a viral pathogen that requires specific strain or strand detection, or a non-coding RNA. Additionally, the RNAscope technology can be combined with immunohistochemistry (IHC) on the same slide for detection of RNA and protein. With catalog probes for over 13,000 targets in 140+ species or the option of made-to-order target probes, the RNAscope technology has now been featured in over 2,000 publications across multiple different research areas including inflammation, immuno-oncology, and virus/pathogen research.

Learn more | acdbio.com/science/applications 17 GMP-grade Immune Cell Culture Reagents

As more and more researchers are investigating the potential of different immune cell populations for therapeutic purposes, the need for innovative tools for ex vivo cell manufacturing has grown. To address this need, we continue to expand our portfolio of GMP-grade reagents, which now includes GMP-grade recombinant proteins along with custom cell separation and CD3/CD28 T cell activation kits manufactured under GMP controls.

Cloudz™ Cell Selection Kits

Bio-Techne is utilizing its pioneering Cloudz dissolvable hydrogel to design a cell selection system that enables the gentle capture and release of select cell populations. Using the principles for cell selection, this technology is ideal for simplifying cell manufacturing protocols, while reducing the risk of bead contamination during downstream cell culture processing. This technology is currently available as a custom service. Features of the technology include: • Size exclusion-based cell selection • Sequential cell selection is available for improved target definition • Manufactured under GMP controls • Compatible with multiple clinical expansion platforms • Customizable for your desired cell population

Principle of the Technology

Step 1: Step 2: Step 3: Cloudz particles bind desired cell Desired cells are retained Cloudz particles are dissolved and desired population cells are collected

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Learn more | rndsystems.com/cloudzselection

18 GMP-grade Recombinant Proteins

R&D Systems GMP-grade recombinant proteins are manufactured under guidelines that allow for their use as ancillary materials in cell therapy or for further manufacturing processes. They come with extensive documentation and traceability, as well as additional quality control testing. Each GMP-grade protein is tested for performance using cell-based bioassays including proliferation, induced cytokine secretion, cytotoxicity, and others. Whenever possible, our GMP proteins are made in an entirely animal-free process using E. coli as the source, although there are some instances when a protein requires production in a eukaryotic system to maintain activity. If this is the case, it is always clearly indicated on the R&D Systems website if a specific GMP-grade protein was not produced in an animal-free process.

GMP-grade Proteins GMP-grade Proteins Molecule Species Source Catalog # Molecule Species Source Catalog # GM-CSF Human E. coli 215-GMP IL-6 Human E. coli 206-GMP IFN-g Human E. coli 285-GMP IL-12 Human Sf 21 (baculovirus) 219-GMP IL-1b Human E. coli 201-GMP IL-15 Human E. coli 247-GMP IL-2 Human E. coli 202-GMP M-CSF Human E. coli 216-GMP IL-3 Human E. coli 203-GMP TGF-b1 Human CHO 240-GMP IL-4 Human E. coli 204-GMP TNF-a Human E. coli 210-GMP

Learn more | rndsystems.com/gmp

GMP Cloudz™ T Cell Activation Kit – CD3/CD28

The Cloudz T Cell Activation Kit – CD3/CD28 contains non-magnetic, dissolvable microparticles that are manufactured under GMP controls and conjugated with the common T cell-activating antibodies, CD3 and CD28. This kit improves T cell expansion and simplifies cell recovery forex vivo manufacturing, while also providing safety and efficacy advantages over current bead-based T cell activation methods. Features of the kit include: • Robust T cell activation and expansion • Simplified recovery of cells – no beads to remove for cell culture • A release buffer that dissolves particles in < 10 seconds

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Learn more | rndsystems.com/cloudzactivation

19 NEW Literature & Interactive Pathways

Chemokine Superfamily

Immune Cell Isolation & Culture Quality Results Start at the Core Wall Poster Bio-Techne Immunoassays — Powered by R&D Systems CD4+ T Cell Subsets

Th2

Th1 Th9

Signal 1 B MHCII TCR-CD3 Complex Th17 Signal 2 Nave CD4+ T Cell APC

Treg Th22

Tfh

Immunology T Y B O O K A C T I V I O M

L A S O Z L E P U Z O R D S S W C R O E O I S R N D G F P I A N G D E O K U S U D S coming soon!

Learn more | rndsystems.com/literature-request

Interactive Pathways Chemokine Signaling Pathways IL-6 Family Signaling Pathways FGF Family Signaling Pathways

Learn more | rndsystems.com/pathways/immunology

Prote294 RnDSy-294 Novus-294 Tocri-294

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