Immune Cell Isolation & Culture

Immune Cell Isolation & Culture Immune Cell Isolation & Culture

Immune cell isolation and culture are necessary for both basic research aimed at investigating the functions of different immune cell types and for expanding specific immune cell populations for therapeutic purposes. Bio-Techne is focused on providing the highest quality reagents for isolating, activating, differentiating, and culturing immune cells. From natural killer (NK) cells and macrophages to dendritic cells and T cells, we offer kits for both cell enrichment and cell differentiation or expansion, along with cell culture media, fetal bovine serum, and an unparalleled selection of recombinant bioactive proteins and antibodies. In addition to our reagents for basic research, we continue to expand our selection of innovative tools for ex vivo cell manufacturing, making it easier for researchers to transition their research from discovery to the clinic. Our portfolio now includes GMP-grade recombinant proteins and GMP-manufactured custom cell separation and CD3/CD28 T cell activation kits, which both utilize non-magnetic Cloudz™ dissolvable microparticle technology.

Contents

Introduction to Immune Cell Isolation & Culture ...... 2 MagCellect™ Cell Selection Kits & Reagents...... 3 T Cell Enrichment Columns...... 4 CellXVivo™ Kits for Immune Cell Differentiation or Expansion...... 4 Cell Culture Media...... 5 Classic Fetal Bovine Serum...... 6 Recombinant Cytokines for Immune Cell Differentiation and Expansion ...... 7 ProDots® Proteins...... 13 Animal-Free™ and Animal Component-Free Process (ACFP) Recombinant Proteins...... 13 GMP-grade Immune Cell Culture Reagents...... 14 Cloudz™ Cell Selection Kits...... 14 GMP-grade Recombinant Proteins...... 15 GMP Cloudz™ T Cell Activation Kit – CD3/CD28...... 15 Custom Services...... 15

Learn more | rndsystems.com/immunecellculture

2 MagCellect™ Cell Selection Kits & Reagents

Isolation or enrichment of a specific cell type of interest from a heterogeneous cell population is frequently a critical first step in immunology research for obtaining a pure population of cells for further characterization. R&D Systems® MagCellect Cell Selection Kits are designed to enrich for specific cell populations based on either a negative or posi- tive selection principle. Features of the kits include:

• Beads with a high binding capacity • Small beads in ferrofluid that have no magnetic memory and will not induce cell damage • Cells can be separated to a very high purity in minutes

MagCellect Human Cell Isolation Kits Catalog # A B Human B Cell Isolation Kit MAGH103 105 105 Human CD14+ Cell Isolation Kit MAGH105 Human CD3+ T Cell Isolation Kit MAGH101 Human Naïve CD4+ T Cell Isolation Kit MAGH115 104 104 Human CD4+ T Cell Isolation Kit MAGH102 Human Memory CD4+ T Cell Isolation Kit MAGH116 CD14 CD14 + + Human CD4 CD25 Regulatory T Cell Isolation Kit MAGH104 103 103 Human CD8+ T Cell Isolation Kit MAGH112

Human Natural Killer Cell Isolation Kit MAGH109 0 0

MagCellect Mouse Cell Isolation Kits Catalog # -103 -103

Mouse B Cell Isolation Kit MAGM204 -103 0 103 104 105 -103 0 103 104 105 Mouse CD3+ T Cell Isolation Kit MAGM201 CD19 CD19 Mouse Naïve CD4+ T Cell Isolation Kit MAGM205 A B Mouse CD4+ T Cell Isolation Kit MAGM202 5 5 Mouse Memory CD4+ T Cell Isolation Kit 10 MAGM206 10 Mouse Naïve CD8+ T Cell Isolation Kit MAGM207 Mouse CD4+CD25+ Regulatory T Cell Isolation Kit MAGM208 104 104 Mouse CD8+ T Cell Isolation Kit MAGM203 Mouse Natural Killer Cell Isolation Kit MAGM210 CD14 CD14 103 103 MagCellect Rat Cell Isolation Kits Catalog # Rat B Cell Isolation Kit MAGR303 0 0 Rat CD3+ T Cell Isolation Kit MAGR301B Rat CD4+ T Cell Isolation Kit MAGR302B -103 -103

-103 0 103 104 105 -103 0 103 104 105 MagCellect Plus Human Kits Catalog # CD19 CD19 Human CD45+ Cell Isolation Kit MAGH125 Isolation of Human CD19+ B Cells from PBMCs using the MagCellect Human B Cell Isolation Kit. Human PBMCs MagCellect Reagents Catalog # were prepared as described in the cell preparation section of the MagCellect Human B Cell Isolation Kit (R&D Systems, MagCellect Magnet MAG997 Catalog # MAGH103). Cells were stained both before (A) MagCellect Streptavidin Ferrofluid MAG999B and after (B) B cell isolation with the MagCellect Kit using a PE-conjugated Mouse Anti-Human CD19 Monoclonal Antibody (R&D Systems, Catalog # FAB4867P) and an APC- conjugated Mouse Anti-Human CD14 Monoclonal Antibody (R&D Systems, Catalog # FAB3832A).

Learn more | rndsystems.com/magcellect

3 T Cell Enrichment Columns

In addition to our MagCellect Kits, Bio-Techne also offers human and mouse T cell enrichment columns from our R&D Systems brand that are designed to enrich for a specific T cell population from a mononuclear cell suspension.

Human T Cell Enrichment Columns Catalog # Mouse T Cell Enrichment Columns Catalog # Human CD3+ T Cell Enrichment Column HTCC Mouse CD3+ T Cell Enrichment Column MTCC Human CD4+ T Cell Enrichment Column HCD4C-1000 Mouse CD4+ T Cell Enrichment Column MCD4C-1000 Human CD4+ T Cell Enrichment Column, Mini HCD43 Mouse CD4+ T Cell Enrichment Column, Mini MCD43 Human CD4+/CD45RA- Memory T Cell Enrichment Column HCD42 Mouse CD8+ T Cell Enrichment Column MCD8C-1000 Human CD4+/CD45RO- Naïve T Cell Enrichment Column HCD41 Human CD8+ T Cell Enrichment Column HCD8C-1000 Human CD8+/CD45RA- Memory T Cell Enrichment Column HCD45 Human CD8+/CD45RO- Naïve T Cell Enrichment Column HCD44

CellXVivo™ Kits for Immune Cell Differentiation or Expansion

R&D Systems CellXVivo Immune Cell Differentiation or Expansion Kits contain optimized combinations of the highest quality proteins and antibodies, along with simple, reproducible protocols for the differentiation or expansion of dendritic cells, macrophages, T helper cell subtypes, regulatory T cells, B cells, natural killer cells, or cytokine-induced killer (CIK) cells. Benefits of the kits include: • Conveniently packages all reagents necessary for immune cell differentiation or expansion • Guaranteed to produce consistent, reliable results • Cost-effective and efficient

A 104 B 104 C 104 D 104 0.9% 0.1% 49.1% 0.1% 0.5% 0.1% 47.6% 0.1%

103 103 103 103

102 102 102 102 IL-17 IL-17 IL-17 IL-17

101 101 101 101

98.5% 0.5% 50.6% 0.1% 99.4% 0.1% 51.9% 0.3% 100 100 100 100 100 101 102 103 104 100 101 102 103 104 100 101 102 103 104 100 101 102 103 104 IL-4 IL-4 IFN-γ IFN-γ Intracellular Cytokine Staining of Mouse Th17 Cells Following Differentiation using the CellXVivo Mouse Th17 Cell Differentiation Kit. Mouse naïve CD4+ T cells without (A, C) or with (B, D) a 5-day differentiation using the reagents provided in the CellXVivo Mouse Th17 Differentiation Kit (R&D Systems, Catalog # CDK017) were stimulated with Cell Activation Cocktail (Tocris, Catalog # 5476) and stained with a PE-conjugated Rat Anti-Mouse IL-17/IL-17A Monoclonal Antibody (Novus Biologicals, Catalog # NBP1-72027) and either an (A, B) APC- conjugated Rabbit Anti-Mouse IL-4 Monoclonal Antibody (Novus Biologicals, Catalog # NBP2-52656) or an (C, D) APC-conjugated Rat Anti-Mouse IFN-g Monoclonal Antibody (R&D Systems, Catalog # IC485A) and detected by flow cytometry. Quadrants were set based on staining with the appropriate isotype controls.

Human CellXVivo Differentiation or Expansion Kits Catalog # Mouse CellXVivo Differentiation Kits Catalog # Human B Cell Expansion Kit CDK005 Mouse Dendritic Cell Differentiation Kit CDK008 Human CIK Expansion Kit CDK014 Mouse Th1 Cell Differentiation Kit CDK018 Human M1 Macrophage Differentiation Kit CDK012 Mouse Th2 Cell Differentiation Kit CDK019 Human M2 Macrophage Differentiation Kit CDK013 Mouse Th17 Cell Differentiation Kit CDK017 Human Monocyte-derived DC Differentiation Kit CDK004 Mouse Treg Cell Differentiation Kit CDK007 Human NK Cell Expansion Kit CDK015 Human Th1 Cell Differentiation Kit CDK001 Human Th2 Cell Differentiation Kit CDK002 Human Th17 Cell Differentiation Kit CDK003C Human Treg Cell Differentiation Kit CDK006

Learn more | rndsystems.com/cellxvivo

4 Cell Culture Media

Bio-Techne now offers R&D Systems 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, our 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 | rndsystems.com/cellculture

5 Classic Fetal Bovine Serum

Fetal Bovine Fetal bovine serum (FBS) is the ideal growth supplement for cell culture media and is generally Serum Premium

Heat-inactivated at C Triple .1 m filtered preferred over other types of sera due to its high levels of nutrients and optimal combination of .2 m filtered Cat. No.: S111 Lot. No.: FBS12 Volume: m growth factors. R&D Systems FBS is produced utilizing strict manufacturing processes and quality Store at -C to -2C For In Vitro Research se Only 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 offers Atlanta Biologicals media supplements, balanced salt solutions, biological buffers, cell freezing media, and more.

Learn more | rndsystems.com/fbs

6 Recombinant Cytokines for Immune Cell Differentiation and Expansion

R&D Systems recombinant and natural proteins are the most referenced proteins in the scientific literature because researchers trust our strong reputation for product quality. We have over 25 years of experience expressing and purifying proteins using a variety of different systems, maintain stringent purification and production standards, and are committed to rigorously testing the bioactivity of our proteins prior to their release to ensure that they will provide researchers with industry-leading bioactivity and lot-to-lot consistency. The section below shows the recombinant proteins and antibodies (when applicable) that are most frequently used for the activation, differentiation, and expansion of different immune cell types, along with specialized serum-free media that has been developed for the culture of human dendritic cells and T lymphocytes. In addition to the standard recombinant proteins shown in the product tables below, our portfolio also includes R&D Systems Animal-Free™ and Animal Component-Free Process recombinant proteins, ProDots® proteins, GMP-grade recombinant proteins, and custom protein development services, which are detailed in the sections that follow.

B Cells Molecule Species Source Catalog # Human CHO 7537-BF BAFF/Blys/TNFSF13B Human NS0 2149-BF Mouse NS0 8876-BF Human HEK293 6420-CL Human E. coli 2706-CL CD40 Ligand/TNFSF5 Human E. coli 6245-CL Mouse CHO 8230-CL Rat CHO 8414-CL Human E. coli 285-IF Human HEK293 10067-IF IFN-g Mouse E. coli 485-MI Rat E. coli 585-IF Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL Human E. coli 204-IL Human CHO 6507-IL IL-4 Mouse E. coli 404-ML Rat E. coli 504-RL Human Sf21 (baculovirus) 217-IL Human Sf21 (stably transfected) 217-ILB IL-10 Human E. coli 1064-IL Mouse E. coli 417-ML Rat E. coli 522-RLB Human E. coli 8879-IL IL-21 Mouse E. coli 594-ML

7 dendritic cell Dendritic Cells Molecule Species Source Catalog # Human Sf21 (baculovirus) 308-FK Flt-3 Ligand Human NS0 308-FKN Mouse NS0 427-FL Human E. coli 215-GM Human CHO 7954-GM GM-CSF Mouse E. coli 415-ML Rat E. coli 518-GM Human E. coli 203-IL IL-3 Mouse E. coli 403-ML Rat E. coli 2524-RL Human E. coli 204-IL Human CHO 6507-IL IL-4 Mouse E. coli 404-ML Rat E. coli 504-RL Human E. coli 288-TPE Human NS0 288-TPN Thrombopoietin Human Sf21 (baculovirus) 288-TP Mouse NS0 488-TO Rat NS0 7864-TP Human E. coli 210-TA Mouse E. coli 410-MT TNF-a Mouse E. coli 410-TRNC Rat E. coli 510-RT

Specialized Media Product Catalog # StemXVivo® Serum-Free Human Dendritic Cell Base Media CCM003

Macrophages Molecule Species Source Catalog # Human E. coli 216-MC M-CSF Human CHO 216-MCC Mouse E. coli 416-ML Human E. coli 285-IF Human HEK293 10067-IF IFN-g Mouse E. coli 485-MI Rat E. coli 585-IF Human E. coli 204-IL Human CHO 6507-IL IL-4 Mouse E. coli 404-ML Rat E. coli 504-RL Human E. coli 213-ILB IL-13 Mouse E. coli 413-ML Rat E. coli 1945-RL

Antibody Application Key B/N Blocking/Neutralization Depl Cell Depletion Ecap ELISA Capture FA Functional Assay FC Flow Cytometry ICC/IF Immunocytochemistry/Immunofluorescence IHC Immunohistochemistry WB Western Blot

8 Myeloid-derived Suppressor Cells (MDSCs) Molecule Species Source Catalog # Human E. coli 214-CS G-CSF Mouse E. coli 414-CS Human E. coli 215-GM Human CHO 7954-GM GM-CSF Mouse E. coli 415-ML Rat E. coli 518-GM Human E. coli 204-IL Human CHO 6507-IL IL-4 Mouse E. coli 404-ML Rat E. coli 504-RL Human E. coli 206-IL Human HEK293 7270-IL IL-6 Mouse E. coli 406-ML Rat E. coli 506-RL Human Sf21 (baculovirus) 217-IL Human Sf21 (stably transfected) 217-ILB IL-10 Human E. coli 1064-IL Mouse E. coli 417-ML Human E. coli 213-ILB IL-13 Mouse E. coli 413-ML cytokine induced killer Rat E. coli 1945-RL Natural Killer (NK) Cells Molecule Species Source Catalog # Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL Human Sf21 (baculovirus) 219-IL Human HEK293 10018-IL IL-12 Mouse Sf21 (baculovirus) 419-ML Mouse NS0 10051-ML Rat Sf21 (baculovirus) 1760-RL Human E. coli 247-ILB IL-15 Mouse E. coli 447-ML Human E. coli 9124-IL Human E. coli B001-5 IL-18/IL-1F4 Mouse E. coli 9139-IL Mouse E. coli B002-5 Human E. coli 8879-IL IL-21 Mouse E. coli 594-ML

Cytokine-induced Killer (CIK) Cells Molecule Species Source Catalog # Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL Human E. coli 285-IF Human HEK293 10067-IF IFN-g Mouse E. coli 485-MI Rat E. coli 585-IF Antibodies Species Clone Catalog # (Applications) Human UCHT1 MAB100 (FA, FC, ICC/IF, IP) CD3 Mouse 145-2C11 MAB484 (Depl., FA, FC, IP)

9 Regulatory T Cells Molecule Species Source Catalog # Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL Human HEK293 7754-BH Human CHO 240-B TGF-b1 Human Platelets 100-B Mouse CHO 7666-MB

Antibodies Species Clone Catalog # (Applications) Human UCHT1 MAB100 (FA, FC, ICC/IF, IP) CD3 Mouse 145-2C11 MAB484 (Depl., FA, FC, IP) Human 37407 MAB342 (FA, WB) CD28 Human Polyclonal AF-342-PB (FA, FC, ICC/IF, IHC) Mouse 794716 MAB4832 (FA, FC) regulatory t cell T Cells Molecule Species Source Catalog # Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL

Specialized Media Product Catalog # StemXVivo® Serum-Free Human T Cell Base Media CCM010

10 Th1 Cells Molecule Species Source Catalog # Human E. coli 285-IF Human HEK293 10067-IF IFN-g Mouse E. coli 485-MI Rat E. coli 585-IF Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL Human Sf21 (baculovirus) 219-IL Human HEK293 10018-IL IL-12 Mouse Sf21 (baculovirus) 419-ML Mouse NS0 10051-ML Rat Sf21 (baculovirus) 1760-RL Human NS0 2526-IL IL-27 Mouse NS0 2799-ML

Th2 Cells Molecule Species Source Catalog # Human E. coli 202-IL IL-2 Mouse E. coli 402-ML Rat E. coli 502-RL Human E. coli 204-IL Human CHO 6507-IL IL-4 Mouse E. coli 404-ML Rat E. coli 504-RL

Antibodies Species Clone Catalog # (Applications) Human UCHT1 MAB100 (FA, FC, ICC/IF, IP) CD3 Mouse 145-2C11 MAB484 (Depl., FA, FC, IP) Human 37407 MAB342 (FA, WB) CD28 Human Polyclonal AF-342-PB (FA, FC, ICC/IF, IHC) Mouse 794716 MAB4832 (FA, FC) Human 25718 MAB285 (B/N, FC, ICC/IF) Human 25718R MAB285R (B/N, ICC/IF) Human K3.53 MAB2852 (B/N, Ecap, FC, WB) Human Polyclonal AF-285-NA (B/N, ICC/IF, WB) IFN-g Human Polyclonal AB-285-NA (B/N, WB) Mouse 37895 MAB485 (B/N, FC, WB) Mouse 37895R MAB485R (B/N, FC) Mouse H22 MAB4851 (B/N, ICC/IF) Mouse Polyclonal AF-485-NA (B/N, WB) Human Polyclonal AF-219-NA (B/N, ICC/IF, WB) IL-12 Mouse Polyclonal AF-419-NA (B/N, WB)

11 Th17 Cells Molecule Species Source Catalog # Human E. coli 201-LB IL-1b/IL-1F2 Mouse E. coli 401-ML Rat E. coli 501-RL Human E. coli 206-IL Human HEK293 7270-IL IL-6 Mouse E. coli 406-ML Rat E. coli 506-RL Human E. coli 8879-IL IL-21 Mouse E. coli 594-ML Human Sf21 (baculovirus) 1290-IL IL-23 Mouse Sf21 (stably transfected) 1887-ML Rat Sf21 (baculovirus) 3136-RL Human HEK293 7754-BH Human CHO 240-B TGF-b1 Human Platelets 100-B Mouse CHO 7666-MB

Antibodies Species Clone Catalog # (Applications) Human UCHT1 MAB100 (FA, FC, ICC/IF, IP) CD3 Mouse 145-2C11 MAB484 (Depl., FA, FC, IP) Human 37407 MAB342 (FA, WB) CD28 Human Polyclonal AF-342-PB (FA, FC, ICC/IF, IHC) Mouse 794716 MAB4832 (FA, FC) Human 25718 MAB285 (B/N, FC, ICC/IF) Human 25718R MAB285R (B/N, ICC/IF) Human K3.53 MAB2852 (B/N, Ecap, FC, WB) Human Polyclonal AF-285-NA (B/N, ICC/IF, WB) IFN-g Human Polyclonal AB-285-NA (B/N, WB) Mouse 37895 MAB485 (B/N, FC, WB) Mouse 37895R MAB485R (B/N, FC) Mouse H22 MAB4851 (B/N, ICC/IF) Mouse Polyclonal AF-485-NA (B/N, WB) Human 34019 MAB204 (B/N, WB) Human 3007 MAB304 (B/N, ICC/IF, WB) Human Polyclonal AF-204-NA (B/N, WB) IL-4 Mouse 30340 MAB404 (B/N, Ecap, WB) Mouse Polyclonal AF-404-NA (B/N, WB) Mouse 30340R MAB404R (B/N, Ecap)

Th17 Cell Markers

To learn about the cell surface and intracellular markers that are most commonly used in the published literature to identify different immune cell types, explore our new Cell Markers Interactive Resource Tool at rndsystems.com/cellmarkers

12 ProDots® Proteins

ProDots® proteins are R&D Systems proteins packaged into easy-to-use lyophilized dots. They are designed to reduce cell culture media preparation time while ensuring optimal protein recovery and bioactivity. Unlike standard lyophilized protein, which is shipped as a diffuse residue in a vial, ProDots proteins are neatly bundled into the shape of a ball so that they can be rolled directly into cell culture media. The benefits of ProDots proteins include: • Eliminates aliquoting • Easy cell culture media preparation • Ensures 100% protein recovery, dissolves instantly • Avoids freeze/thaw cycles

ProDots Recombinant Human Proteins ProDots Recombinant Human Proteins Molecule Catalog # Molecule Catalog # Flt-3 Ligand PRD308 IL-3 PRD203 G-CSF PRD214 IL-4 PRD204 GM-CSF PRD215 IL-6 PRD206 IFN-g PRD285 IL-10 PRD217 IL-1b PRD201 M-CSF PRD216 IL-2 PRD202 TGF-b1 PRD240

For a complete, up-to-date product listing | rndsystems.com/prodots

Animal-Free™ and Animal Component-Free Process (ACFP) Recombinant Proteins

Animal-free proteins are particularly important for researchers concerned with experimental variables caused by trace animal components or mammalian pathogens. To support developing research and bioproduction demands, Bio-techne offers R&D Systems Animal-Free recombinant proteins, which are manufactured in a laboratory exclusively dedicated to the production and purification of proteins under animal-free conditions. We also offer R&D Systems Animal Component-Free Process (ACFP) recombinant proteins that are expressed in an animal-free certified Sf 9 insect cell line using dedicated animal-free raw materials and labware. Features of our Animal-Free proteins include: • Endotoxin Levels < 1.0 EU per 1 µg of product • Tested to ensure no impairment of biological activity • Additional documentation can be provided for ex vivo research or bioproduction

Animal-Free or Animal Component-Free Process Proteins Animal-Free or Animal Component-Free Process Proteins Molecule Species Source Catalog # Molecule Species Source Catalog # CD40 Ligand/TNFSF5 Human E. coli AFL617 IL-2 Human E. coli AFL202 Flt-3 Ligand Human Sf9 (baculovirus) ACFP308 IL-3 Human E. coli AFL203 G-CSF Human E. coli AFL214 IL-4 Human E. coli AFL204 Human E. coli AFL215 IL-6 Human E. coli AFL206 GM-CSF Mouse E. coli AFL415 Human E. coli AFL216 M-CSF IFN-g Human E. coli AFL285 Mouse E. coli AFL416 Human E. coli AFL201 Human E. coli AFL210 IL-1b/IL-1F2 TNF-a Mouse E. coli AFL401 Mouse E. coli AFL410

For a complete, up-to-date product listing | rndsystems.com/animalfree

13 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 population desired cells are collected

Desired cell bound to Desired cells Unanted cell Cell strainer antibody conugated Cloud particle

Unanted cells

Learn more | rndsystems.com/cloudzselection

14 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. The R&D Systems website always clearly indicates whether a specific GMP-grade protein is produced in an animal-free process or not.

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

Cloud CD3CD28 T Cells or BC CloudCell Complex T Cell Expansion icrosphere-free T Cells Donstream Applications

25 Add elease Buffer 20 15

+ Cell umber elative Cloud dissolve 5 1 seconds

0 100 101 102 103 104 CD4 Learn more | rndsystems.com/cloudzactivation

Custom Services

If you still don’t see what you need, let us know. Bio-Techne offers expert custom cell culture media production services along with custom protein and antibody services to ensure that we can meet your research needs. Learn more | rndsystems.com/custom

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