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Luminex® Quarterly Fall 2019

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Quality Results Start at the Core .................................................................3What’s New! R&D Systems® Luminex® Assays ...........................................3Instruments ..................................................................................................3Technical Application Note ..........................................................................4Troubleshooting Guide .................................................................................7Luminex Assay Customization Tool .......................................................... 14Custom Luminex Services ........................................................................ 15Which Immunoassay Is Right For You? .................................................... 16

Table of Contents

3Learn more | rndsystems.com/luminex-assays-and-high-performance-assays

Quality Results Start at the Core

What’s New! R&D Systems® Luminex® Assays

Instruments

Your results matter. So, what’s inside your immunoassay should too. R&D Systems’ antibodies and proteins are the core of every Bio-Techne immunoassay platform. Assays that are backed by over 68,000 citations and the expertise behind our gold-standard Quantikine® ELISAs. Antibodies are highly specifi c, manufactured in-house to ensure reproducibility and tested for suitability in selected applications.

Have an existing Luminex analyzer already? R&D Systems Luminex Assays are compatible with all Luminex instrumentation systems. These systems can be obtained from R&D Systems.

Learn more @ https://www.rndsystems.com/products/luminex-instrumentation

Luminex MAGPIX Luminex 100/200 Luminex FLEXMAP 3D

Human Th1/Th2 Fixed

Human Th9 / Th17 / Th22 FixedHuman Immunotherapy Fixed

Human Cytokine Fixed

Human Chemokine Fixed

Human Growth Factor Fixed

Human XL Cytokine Fixed

What’s at the core of your immunoassay?

New Performance Panels available off-the-shelf and ready to ship, allowing you to make your discoveries sooner.

NHP XL Discovery Performance Assay - Coming Soon

Human Metabolic Performance Assay - Coming Soon

LuminexFixed PanelsT Cells | Immunotherapy | Cytokines | Chemokines

4 Learn more | rndsystems.com/luminex

Technical Application Note

Luminex® Multiplex Immunoassays for Classifi cation of Th1, Th2, and Th17 CD4+

T Cell Differentiation

Developing CD4+ T cells in the thymus differentiate into various distinct subsets. Each subset produces a characteristic combination of cytokines to activate other immune cells, control ongoing immune responses, or manage immune memory.

Bio-Techne has developed R&D Systems® Luminex®

Immunoassays to specifi cally quantify biomarkers of Th1/Th2 and Th9/Th17/Th22 differentiated CD4+

T cells. These two Luminex kits allow for the characterization of T cell lineages present in CD4+

T cells which have been induced towards specifi cT cell subsets.

METHODS

Cell Culture

Using the MagCellectTM Human CD4+T cell Isolation Kit (R&D Systems; Catalog # MAGH102), CD4+T cells were isolated from Human peripheral blood mononuclear cells cultured in RPMI and supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin sulfate. These cells were either untreated or treated with Th1, Th2 or Th17-inducing reagents.

Th1 Cells: Flasks were coated with 2 µg/mL goat anti-mouse IgG (AF007) overnight at room temperature. Wash 1X with PBS, and then coat with 1 µg/mL Ms x hCD3ε (MAB100) 2-3 hours at room temperature. Wash 1X with PBS prior to adding CD4+T cells seeded at 0.5 – 1 x 106/mL. To activate the cells the following polarizing cytokines and antibodies were then added to the fl ask and incubated for 5 days:

20 ng/ml rhIL-2 (202-IL)40 ng/ml rhIL-12 (219-IL)

5 µg/ml Ms x hIL-4 (MAB304)5 µg/ml Ms x hCD28 (MAB342)

Further incubate stimulated cells for 24 hours with PMA (10 ng/mL; Tocris; Catalog # 1201/1) and Ionomycin calcium salt (500 ng/mL; Tocris; Catalog # 1704/1)

Th17 cells: Flasks were coated with 2 µg/mL goat anti-mouse IgG (AF007) overnight at room temperature. Wash 1X with PBS, and then coat with 1 µg/mL Ms x hCD3ε (MAB100) 2-3 hours at room temperature. Wash 1X with PBS prior to adding CD4+T cells seeded at 0.5 – 1 x 10e6/mL. To activate the CD4+ T cells the following cytokines and antibodies were then added to the fl ask and incubated for 5 days:

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RESULTS Cells polarized towards a Th1 phenotype secreted increased levels of IFN-a and TNF-b, as expected for Th1 differen-tiation (Figure 1B). A hallmark of Th2 differentiation is increased levels of IL-4, IL-5, and IL-13, which we observed (Figure 1A). Stimulating cells with IL-2, IL-23, IL-1b and IL-6 drove activated CD4+ T cell differentiation to a Th17 phenotype, as observed by the secretion of IL-17A (Figure 1D).

CONCLUSION The cytokine profiles measured with our Luminex Fixed Panels for Th1/Th2 and Th9/Th17/Th22 were consistent with expected results for each respective T cell phenotype. R&D Systems fixed panels provide a convenient, efficient, and highly-validated immunoassay option to generate biomarker/cytokine profiles in response to intracellular and extracellular pathogenic bacteria, viruses, and fungi. These fixed panels are provided as an off-the-shelf product, making it easier and faster than ever to get the Luminex kits you need.

Further incubate stimulated cells for 24 hours with PMA (10 ng/mL; Tocris; Catalog # 1201/1) and Ionomycin calcium salt (500 ng/mL; Tocris; Catalog # 1704/1) Th2 cells: Add CD4+T cells seeded at 0.5 – 1 x 10e6/mL to an un-coated flask. To activate the CD4+ T cells the following were then added to the flask and incubated for 5 days:

Further incubate stimulated cells for 24 hours with PMA (10 ng/mL; Tocris; Catalog # 1201/1) and Ionomycin calcium salt (500 ng/mL; Tocris; Catalog # 1704/1) Luminex Fixed Panel Analysis Cell culture supernates were analyzed with either the Luminex Th1/Th2 Fixed Panel (Catalog # LKTM008) or the Luminex Th9/Th17/Th22 Fixed Panel (Catalog # LKTM009). Samples were assayed according to the procedures outlined in the product inserts. Both kits are available from R&D Systems.

20 ng/mL rhIL-2 (202-IL) 20 ng/mL rhIL-23 (1290-IL) 10 ng/mL rhIL-1β (201-LB)

20 ng/ml rhIL-2 (202-IL)

40 ng/mL rhIL-6 (206-IL) 5 µg/mL Ms x hCD28 (MAB342)

5 ug/ml PHA

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Figure 1 - Th1, Th2 and Th17 Cytokine Differentiation Profiles A & B - Cytokine profile for the TH1/Th2 fixed panel analytes. C & D - Cytokine profile for the Th9/Th17/Th22 fixed panel anayltes. + = values above the limits of the standard curve * = stimulating cytokine aCD3/28 = cells from an uncoated flask incubated in RPMI and 10% FBS for 5 days NT = Non-treated cells from an antibody coated flask with only Ms x hCD28 added for 5 days Note: The concentrations represented in this figure have not been corrected for the initial 1:2 sample dilution

0

A

+

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GM-CSF IFN-γ TNF-α

+ +

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+ + +

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IL-12 p70 IL-13 IL-1

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Th2

IL-4 IL-50

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Analyte

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IL-10 IL-12p70

IL-13 IL-15 IL-17E IL-33 IL-4 IL-5

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CD40 Ligand IFN-γ IL-17A MIP-3α TNF-α

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Th17

+ +

+ +

REFERENCES 1. Suntharalingam, G. et al. (2006) N. Engl. J. Med. 335:1018. 2. Maude, S.L. et al. (2014) Cancer J. 20:119. 3. Römer, P.S. et al. (2011) Blood 118:6772. 4. Findlay, L. et al. (2010) J. Immunol. Methods 352:1.

5. Stebbings, R. et al. (2013) J. Immunotoxicol. 10:75. 6. Attarwala, H. (2010). J. Young Pharm. 2:3 7. Hünig, T. et al. (2011) Adv Immunol. 95:111.

ORDERING INFORMATION

Fixed Panels Max Number Analytes Catalog #Human Th1/Th2 Discovery 11 LKTM008

Human Th9/Th17/Th22 Discovery 17 LKTM009

Customizable Panels Number Analytes to Select Catalog #Human XL Cytokine Discovery (Premixed) 45 FCSTM18

Human XL Cytokine Discovery (User mixed) 45 LUXLM000

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Troubleshooting Guide

Observation Possible Source SuggestionAcquisition Problems and Error Messages

Incompatible instrument Instrument is out of calibration Incorrect probe height Sample probe is clogged Microparticle spectral address is not assigned correctly Incorrect instrument settings

Use the instrument that is compatible to the microparticle type. R&D Systems® Luminex® assays are compatible with all Luminex® instruments. • Luminex® 100/200™ • FLEXMAP 3D™ • Luminex® MAGPIX®

To obtain accurate measurements, regular calibration of the instrument is required. Best practice is to run assays within one week of calibration. Luminex® recommends running verification the day of the assay to confirm the instrument is functioning properly and is within current calibration settings. Perform instrument calibration and verification per the instrument user’s manual. Adjust the sample probe vertical height and align to the plate per the instrument user’s manual. Clean the sample probe per the instrument user’s manual. Replace the sample probe if necessary. Remember to readjust the vertical height each time the probe has been removed. Ensure microparticle regions are assigned correctly per the kit insert or the Certificate of Analysis. Microparticle maps are custom created depending upon the analytes selected. In the event of omission or an incorrect assignment of a microparticle region, data will be missing in the CSV file, try the “Replay” function to retrieve the data following selection of the appropriate microparticle regions. Follow the insert instructions on instrument settings.

Low Microparticle Count Instrument is out of calibration Wrong event or microparticle setting The system is timed-out (Luminex® 100/200™ and FLEXMAP3D™) Sample contains debris which affects acquisition Miscalculation of microparticle dilution/lower number of microparticles added per well Microparticles are clumped or aggregated Microparticles not in suspension during acquisition Shaker with incorrect settings Magnetic microparticles not collected at the bottom of plate during wash steps Sample was run undiluted Blockage of sample probe

Perform instrument calibration and verification. To obtain accurate measurements regular calibration of the instrument is required. Best practice is to run assays within one week of calibration. Luminex® recommend running verification the day of the assay to confirm the instrument is functioning properly with current calibration settings. Verify that the events/bead is set at 50. A 50-count per analyte is sufficient to produce a statistically accurate result. A bead count of about 25 may be acceptable if the samples are run in duplicates and other parameters of measuring the performance of the assay are fine. If your instrument times out when using flow cytometry-based instruments such as the Luminex® 100/200™, stop the plate run. Check the probe height and confirm the magnetic microparticle type is selected. Then re-run the plate. As each microparticle has a different rate for acquisition and the instrument is set to collect 50 microparticles in a designated time, a “time-out” may result in insufficient microparticle counts for one or more analytes. The MagPIX® instrument has a fixed read time for each well and does not have time-out functionality. Centrifuge samples on the day of the assay at approximately 16,000 x g for 4 minutes immediately before use. In rare cases, an extended centrifugation may be necessary. Confirm microparticles were diluted according to the kit insert. Centrifuge the microparticle cocktail concentrate (for 30 seconds at 1,000xg) and gently vortex the concen-trated before preparing the 1X diluted microparticle cocktail. Immediately before placing the plate on the reader, shake the plate for one additional minute in 1X Wash Buffer to resuspend the microparticles. Use a horizontal orbital microplate shaker with a 0.12” orbit. Ensure the shaker speed is set per recommenda-tions from the kit insert. Use an appropriate magnetic device designed to accommodate a microplate. Wash by applying the magnet to the bottom of the microplate, allow 1 minute before decanting wash buffer. Do not blot dry as this may cause a loss of microparticles. Samples require at least a 2-fold dilution with the appropriate Calibrator Diluent. Mix thoroughly. Samples may require higher than 2-fold dilutions. Review the Product Insert, Certificate of Analysis or R&D Systems® Luminex® Assay Customization Tool for the suggested starting dilution for each sample type. See above under acquisition and error messages.

Low Fluorescence Intensity (FI) signal or Poor Sensitivity

Non-optimal preparation of the standard curve Non-optimal dilution of the detection antibodies or streptavidin-PE concentrates Photo-bleaching of the PE signal Incorrect shaker settings Incorrect instrument settings

Confirm the standard reconstitution volume from the standard value card or the Certificate of Analysis. Incorrect reconstitution of the standard will result in inaccurate sample value calculations. Be sure to follow reconstitution instructions for all lyophilized reagents outlined in the kit insert. Confirm reagent dilutions were performed according to the kit insert. Streptavidin-PE is light sensitive. Protect from light. See above. See above.

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Observation Possible Source SuggestionSample Readings are Out of Range (OOR Error Message)

Samples are below assay range (<OOR Error Messages) and contain no analyte, or the analyte level is below the level of detection, or the sample may be too diluted. When readings are above assay range (>OOR Error Messages)

Analyte of interest may be undetectable in the assay range due to low abundance of natural protein. Check kit instructions or the R&D Systems® Luminex® Assay Customization Tool for suggested sample dilution. Suggested dilution factors are based on samples from healthy volunteers. Depending on the unique nature of an individual sample, a different dilution factor may be needed to bring the reading within the dynamic range of the assay. Review the Product Insert, Certificate of Analysis or Check kit instructions or the R&D Systems® Luminex® Assay Customization Tool for the suggested initial dilution. Suggested dilution factors are based on samples from healthy volunteers. Depending on the nature of the sample, it may require a further dilution to bring the reading within the assay range. Note - Samples may require dilution and re-analysis if a specific analyte is out of range.

Poor Precision with sample measurements

Presence of interfering components in samples, especially samples with complex matrices such as plasma and serum Sample type not validated for the assay Samples with hemolyzed and hyperlipidemic matrices Integrity of the sample is compro-mised while in storage Non-optimal pipetting technique Assay reagents not equilibrated to room temperature prior to use

Check for the presence of interfering components, additives, or if gel separators were introduced into the sample by performing a Spike/Recovery and Linearity test. Contact R&D Systems® Technical Service if you require assistance with this test. Check the kit insert to confirm if the sample type has been validated for the assay. Avoid the use of samples with hemolyzed or hyperlipidemic matrices. Such samples may disrupt antibody binding or clog the probe. See discussion above on how to clean the sample probe. Follow the kit insert on Sample Collection & Storage. Observe best practices for processing and storing the samples after collection. Avoid repeated freeze-thaw cycles. Ensure a consistent and accurate pipetting method. Dispense microparticles, diluents and samples accurately. Change pipette tips between samples and dilutions. Pre-wet tips for sample replicates. Ensure that your pipettes are calibrated regularly. All assay components should be equilibrated to room temperature prior to use.

High Background Signal Incorrect buffer used for the dilution of standards and/or samples Blank wells accidently spiked with standard or samples Extended incubation with detection Abs or streptavidin-PE

Ensure the use of the recommended Calibrator Diluent for the dilution of standards/samples per kit insert. Do not add standard or samples to wells designated as blank. Add Calibrator Diluent only. Follow the kit instructions for incubation times and follow precisely.

Microparticle Aggregation Samples with hemolyzed and hyperlipidemic matrices Microparticles not thoroughly mixed Doublet Discriminator gates setting is incorrect

See above Follow the kit instructions on the preparation of the diluted microparticle cocktail. Use a plate shaker with appropriate settings for the assay. Shake plate for one additional minute in 1X Wash Buffer immediately before analysing in an appropriate instrument. Check the kit insert for the Doublet Discriminator gate settings and adjust settings as needed.

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Luminex® Menu

Angiogenesis Cat. #s FCSTM02/LAMN000• Angiogenin• Angiopoietin-1• Endostatin

• FGF acidic• FGF basic• PDGF-AA

• PDGF-BB• PlGF• Thrombospondin-2

• VEGF• VEGF-D

Biomarker Custom Cat. #s FCSTM13/LBAM000• BAFF/BLyS/TNFSF13B• CCL20/MIP-3 alpha• CD14

• CD25/IL-2 R alpha• CD27/TNFRSF7• CXCL13/BLC/BCA-1

• gp130• IL-6 R alpha• TNF RII/TNFRSF1B

Cardiac A Custom Cat. #s FCSTM11/LUCAM000• CD40 Ligand/TNFSF5 • GDF-15

• Pappalysin-1/PAPP-A• Proprotein Convertase 9/PCSK9

• ST2/IL-33 R• TNF RII/TNFRSF1B

Cardiac B Custom Cat. #s FCSTM12/LUCBM000• C-Reactive Protein/CRP• Cystatin C

• Myeloperoxidase/MPO• P-Selectin/CD62P

• Serpin E1/PAI-1• TIMP-1

Cytokine A Custom Cat. #s FCSTM03/LUHM000• CCL2/JE/MCP-1• CCL3/MIP-1 alpha• CCL4/MIP-1 beta• CCL5/RANTES• CXCL5/ENA-78• FGF basic

• G-CSF• GM-CSF• IFN-gamma• IL-1 alpha/IL-1F1• IL-1 beta/IL-1F2• IL-10

• IL-17/IL-17A• IL-1ra/IL-1F3• IL-2• IL-4• IL-5• IL-6

• IL-8/CXCL8• Thrombopoietin/Tpo• TNF-alpha• VEGF

Cytokine B Custom Cat. #s FCSTM04/LUBM000• CCL11/Eotaxin• CD40 Ligand/TNFSF5 • CXCL10/IP-10/CRG-2

• CXCL11/I-TAC• EGF• HGF

• IL-12 p70 IL-13• Leptin/OB

Chemokine Fixed Cat. # LKTM012• CCL11/Eotaxin• CCL2/JE/MCP-1

• CCL3/MIP-1 alpha• CCL4/MIP-1 beta

• CCL5/RANTES• CXCL1/GRO alpha/KC/CINC-1

• CXCL10/IP-10/CRG-2• IL-8/CXCL8

Cytokine Fixed Cat. # LKTM011• IFN-alpha• IFN-gamma• IL-1 alpha/IL-1F1• IL-1 beta/IL-1F2

• IL-1ra/IL-1F3• IL-2• IL-3 • IL-4

• IL-6 • IL-7• IL-10• IL-15

• IL-33• VEGF

Growth Factor Fixed Cat. # LKTM013• CCL3/MIP-1 alpha• CCL19/MIP-3 beta• CD40 Ligand/TNFSF5• CX3CL1/Fractalkine• CXCL2/GRO beta/MIP-2/CINC-3• EGF

• FGF basic/FGF2• Flt-3 Ligand• G-CSF• Granzyme B• IFN-beta• IL-3

• IL-8/CXCL8• IL-17E/IL-25• IL-33• PD-L1/B7-H1• PDGF-AA• PDGF-AB/BB

• TGF-alpha• TRAIL/TNFSF10• VEGF

Immunotherapy Fixed Cat. # LKTM010• CCL2/JE/MCP-1• CCL3/MIP-1 alpha• CCL4/MIP-1 beta• CD40 Ligand/TNFSF5• CXCL10/IP-10/CRG-2• GM-CSF

• Granzyme B• IFN-alpha• IFN-gamma• IL-1 alpha/IL-1F1• IL-1 beta/IL-1F2• IL-1ra/IL-1F3

• IL-2• IL-4• IL-6• IL-8/CXCL8• IL-10• IL-12 p70

• IL-13• IL-15• IL-17/IL-17A• IL-33• PD-L1/B7-H1• TNF-alpha

Adhesion Fixed Cat. # LKTM007• E-Selectin/CD62E • P-Selectin/CD62P • ICAM-1/CD54 • VCAM-1/CD106

Human High Performance Panels

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Th1/Th2 Fixed Cat. # LKTM008• GM-CSF• IFN-gamma• IL-1 beta/IL-1F2

• IL-2• IL-4• IL-5

• IL-6• IL-10• IL-12 p70

• IL-13• TNF-alpha

Th9/Th17/Th22 Fixed Cat. # LKTM009• CCL20/MIP-3 alpha• CD40 Ligand/TNFSF5• GM-CSF• IFN-gamma• IL-1 beta/IL-1F2

• IL-2• IL-4• IL-5• IL-6• IL-10

• IL-12 p70• IL-13• IL-15• IL-17/IL-17A• IL-17E/IL-25

• IL-33• TNF-alpha

TIMP Fixed Cat. # LKTM003• TIMP-1 • TIMP-2 • TIMP-3 • TIMP-4

High Sensitivity Cytokine Panel A CustomCat. # FCSTM09/ LHSCM000• CXCL8/IL-8• GM-CSF• IFN-gamma

• IL-1 beta/IL-1F2• IL-2• IL-4

• IL-5• IL-6• IL-10

• IL-12 p70• TNF-alpha• VEGF

High Sensitivity Cytokine Panel B CustomCat. # FCSTM14/LBHS000• GM-CSF• IFN-gamma• IL-1 beta/IL-1F2• IL-2• IL-5

• IL-6• IL-7• IL-13• IL-15• IL-17A

• IL-17F• IL-22• IL-23• IL-31• IL-33

• IL-36 beta• TNF-alpha

Kidney Biomarker Panels Custom Cat. #s FCSTM16/LHK000• Clusterin• CXCL10/IP-10/CRG-2• Cystatin C

• Lipocalin-2/NGAL• Osteopontin/OPN• RBP4/Retinol-Binding Protein 4

• TFF3• TIM-1/KIM-1/HAVCR

MMP Custom Panels Cat. #s FCSTM07/LMPM000• EMMPRIN/CD147• MMP-1• MMP-10

• MMP-12• MMP-13• MMP-2

• MMP-3• MMP-7• MMP-8

• MMP-9

Obesity Custom Panels Cat. #s FCSTM10/ LOBM000• Adiponectin/Acrp30• CCL2/JE/MCP-1• Complement Factor D/Adipsin

• C-Reactive Protein/CRP• IL-10• IL-6

• Leptin/OB• Resistin• Serpin E1/PAI-1

• TNF-alpha

TGF-beta Custom Panels Cat. #s FCSTM11/ LTGM000• TGF-beta 1 • TGF-beta 2 • TGF-beta 3

XL Cytokine Fixed Panels Custom Cat. #s FCSTM18/ LUXLM000 Fixed Cat. # LKTM014• CCL2/JE/MCP-1• CCL3/MIP-1 alpha• CCL4/MIP-1 beta• CCL5/RANTES• CCL11/Eotaxin• CCL19/MIP-3 beta• CCL20/MIP-3 alpha• CD40 Ligand/TNFSF5• CX3CL1/Fractalkine• CXCL1/GRO alpha/KC/CINC-1• CXCL2/GRO beta/MIP-2/CINC-3• CXCL10/IP-10/CRG-2

• EGF• FGF basic/FGF2• Flt-3 Ligand• G-CSF• GM-CSF• Granzyme B• IFN-alpha 2• IFN-beta• IFN-gamma• IL-1 alpha/IL-1F1• IL-1 beta/IL-1F2• IL-1ra/IL-1F3

• 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-alpha• TNF-alpha• TRAIL/TNFSF10• VEGF

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ADAMTS13 CCL4/MIP-1 beta

Adiponectin/Acrp30 CCL5/RANTES

Aggrecan CCL7/MCP-3/MARC

AgRP/ART CCL8/MCP-2

ALCAM/CD166 CD117/c-kit

Aldehyde Dehydrogenase 1-A1/ALDH1A1

CD14

alpha 1-Microglobulin CD163

alpha 2-Macroglobulin CD23/Fc epsilon RII

alpha-Fetoprotein/AFP CD25/IL-2 R alpha

alpha-N-acetylglucosaminidase/NAGLU CD27/TNFRSF7

alpha-Synuclein CD30/TNFRSF8

Angiogenin CD31/PECAM-1

Angiopoietin-1 CD40 Ligand/TNFSF5

Angiopoietin-2 CD40/TNFRSF5

Angiopoietin-like Protein 3/ANGPTL3 CD44

Angiopoietin-like Protein 4/ANGPTL4 CEACAM-1/CD66a

Angiopoietin-like Protein 6/ANGPTL6 CEACAM-5/CD66e

APP Chemerin

APRIL/TNFSF13 Chitinase 3-like 1

Atrial Natriuretic Peptide/ANP Coagulation Factor III/Tissue Factor

B7-H3 Coagulation Factor XIV/Protein C

BAFF/BLyS/TNFSF13B Collagen I alpha 1

BCMA/TNFRSF17 Collagen IV alpha 1

BDNF Complement Component C2

beta 2-Microglobulin Complement Component C5a

beta-NGF Complement Component C9

BMP-10 Complement Factor D/Adipsin

BMP-2 Contactin-1

BMP-4 C-Peptide

BMP-7 C-Reactive Protein/CRP

BMP-9 Cripto

CA125/MUC16 CTRP9/C1qTNF9

CA15-3/MUC-1 CX3CL1/Fractalkine

Calbindin D CXCL1/GRO alpha/KC/CINC-1

Carbonic Anhydrase IX/CA9 CXCL10/IP-10/CRG-2

Cathepsin S CXCL11/I-TAC

CCL1/I-309/TCA-3 CXCL12/SDF-1

CCL11/Eotaxin CXCL13/BLC/BCA-1

CCL13/MCP-4 CXCL14/BRAK

CCL14/HCC-1/HCC-3 CXCL16

CCL15/MIP-1 delta CXCL2/GRO beta/MIP-2/CINC-3

CCL17/TARC CXCL4/PF4

CCL18/PARC CXCL5/ENA-78

CCL19/MIP-3 beta CXCL6/GCP-2

CCL2/JE/MCP-1 CXCL7/NAP-2

CCL20/MIP-3 alpha CXCL9/MIG

CCL21/6Ckine Cystatin C

CCL22/MDC DcR3/TNFRSF6B

CCL23/MPIF-1 D-dimer

CCL24/Eotaxin-2/MPIF-2 Dkk-1

CCL25/TECK DPPIV/CD26

CCL26/Eotaxin-3 DR3/TNFRSF25

CCL27/CTACK EGF

CCL28 EMMPRIN/CD147

CCL3/MIP-1 alpha Endocan/ESM-1

Endoglin/CD105 IGFBP-3

Endostatin IGFBP-4

Endothelin-1 IGFBP-6

Enolase 2/Neuron-specific Enolase IGFBP-rp1/IGFBP-7

ENPP-2/Autotaxin IL-1 alpha/IL-1F1

EN-RAGE/S100A12 IL-1 beta/IL-1F2

EpCAM/TROP1 IL-1 RI

EphA2 IL-1 RII

ErbB2/Her2 IL-10

ErbB3/Her3 IL-11

ESAM IL-12 p70

E-Selectin/CD62E IL-12/IL-23 p40

FABP4/A-FABP IL-13

Fas Ligand/TNFSF6 IL-15

Fas/TNFRSF6/CD95 IL-16

Ferritin IL-17/IL-17A

Fetuin A/AHSG IL-17C

FGF acidic IL-17E/IL-25

FGF basic IL-18/IL-1F4

FGF-13 IL-19

FGF-23 IL-1ra/IL-1F3

Fibroblast Activation Protein alpha/FAP IL-2

Fibronectin IL-21

Flt-3 Ligand IL-22

Follistatin-like 1/FSTL1 IL-23

Follistatin-related Gene Protein/FLRG IL-27

FRSF9/CD137 IL-28A/IFN-lambda 2

Furin IL-28B/IFN-lambda 3

Galectin-1 IL-3

Galectin-3 IL-31

Galectin-3BP/MAC-2BP IL-33

Galectin-9 IL-34

Gas6 IL-36 beta/IL-1F8

G-CSF IL-4

GDF-15 IL-4 R alpha

GDNF IL-5

GITR/TNFRSF18 IL-6

GM-CSF IL-6 R alpha

gp130 IL-7

Granzyme A IL-8/CXCL8

Granzyme B Insulin

Growth Hormone ITIH4

HB-EGF Kallikrein 3/PSA

HE4/WFDC2 Kallikrein 5

HGF Kallikrein 6/Neurosin

HGF R/c-MET Lactoferrin

HMW Adiponectin/Acrp30 LBP

HTRA2/Omi Leptin R

ICAM-1/CD54 Leptin/OB

IFN-alpha LIF

Human Luminex Assay, Analyte MenuCat. # LXSAHMHighlighted analytes are unique to R&D Systems

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IFN-beta LIGHT/TNFSF14

IFN-gamma Lipocalin-2/NGAL

IFN-gamma R1/CD119 LRG1

IGFBP-1 L-Selectin/CD62L

IGFBP-2 Lumican

Lymphotoxin-alpha/TNF-beta Proprotein Convertase 9/PCSK9

MAdCAM-1 Protein S/PROS1

MBL Proteinase 3/Myeloblastin/PRTN3

MCAM/CD146 P-Selectin/CD62P

M-CSF RAGE/AGER

M-CSF R/CD115 RBP4/Retinol-Binding Protein 4

Mesothelin Reg3A

MFG-E8 Relaxin-2

MIA Renin

MICA Resistin

MICB ROBO4

Midkine S100A8

MIF S100A9

MMP-1 S100B

MMP-10 SCF/c-kit Ligand

MMP-12 SCGF/CLEC11a

MMP-13 Serpin A10/ZPI

MMP-2 Serpin A12

MMP-3 Serpin A4/Kallistatin

MMP-7 Serpin A7/TBG

MMP-8 Serpin B3/SCCA1

MMP-9 Serpin C1/Antithrombin-III

MSP/MST1 Serpin E1/PAI-1

Myeloperoxidase/MPO Serpin F1/PEDF

Myoglobin SHBG

N-Cadherin SLPI

NCAM-1/CD56 SOST/Sclerostin

Nectin-4 SPARC

Nephrin SP-D

Neuregulin-1 beta 1/NRG1 beta 1 ST2/IL-33 R

Neuropilin-1 Stanniocalcin 1/STC-1

NT-3 Syndecan-1/CD138

NT-4 Syndecan-4

Oncostatin M/OSM TACI/TNFRSF13B

Osteoactivin/GPNMB Tau

Osteopontin/OPN Tenascin C

Osteoprotegerin/TNFRSF11B TFF3

Park7/DJ-1 TFPI

PBEF/Visfatin TfR (Transferrin R)

PDGF-AA TGF-alpha

PDGF-AB Thrombomodulin/BDCA-3

PDGF-BB Thrombopoietin/Tpo

PDGF-CC Thrombospondin-2

PDGF-DD Thymidine Kinase 1

PD-L1/B7-H1 Tie-2

Pentraxin 2/SAP TIM-1/KIM-1/HAVCR

Pentraxin 3/TSG-14 TIMP-1

Periostin/OSF-2 TNF RI/TNFRSF1A

PLA2G7/PAF-AH/Lp-PLA2 TNF RII/TNFRSF1B

PlGF TNF-alpha

PP14/Glycodelin Total Inhibin

Procalcitonin TRACP/PAP/ACP5

Progranulin/PGRN TRAIL R2/TNFRSF10B

Prolactin TRAIL R3/TNFRSF10C

Properdin TRAIL/TNFSF10

TRANCE/TNFSF11/RANK L Uteroglobin/SCGB1A1

TREM-1 VAP-1/AOC3

TSLP VCAM-1/CD106

UCH-L1/PGP9.5 VEGF

ULBP-1 VEGF R1/Flt-1

ULBP-2/5/6 VEGF R2/KDR/Flk-1

ULBP-3 VEGF R3/Flt-4

ULBP-4/RAET1E VEGF-C

uPAR VEGF-D

u-Plasminogen Activator (uPA)/Urokinase

vWF-A2

Uromodulin XCL1/Lymphotactin

Mouse Luminex Assay, Analyte Menu# LXSAMSMHighlighted analytes are unique to R&D Systems

Active Ms Analytes GDF-15

Adiponectin/Acrp30 GM-CSF

Angiopoietin-2 Granzyme B

BAFF/BLyS/TNFSF13B Haptoglobin

beta-NGF HGF

C1q R1/CD93 ICAM-1/CD54

CCL11/Eotaxin IFN-gamma

CCL12/MCP-5 IGFBP-1

CCL19/MIP-3 beta IGFBP-3

CCL2/JE/MCP-1 IGF-I/IGF-1

CCL20/MIP-3 alpha IL-1 alpha/IL-1F1

CCL21/6Ckine IL-1 beta/IL-1F2

CCL22/MDC IL-10

CCL3/MIP-1 alpha IL-12 p70

CCL4/MIP-1 beta IL-13

CCL5/RANTES IL-16

CCL7/MCP-3/MARC IL-17/IL-17A

CCL8/MCP-2 IL-17E/IL-25

Chitinase 3-like 1 IL-2

C-Reactive Protein/CRP IL-27

CXCL1/GRO alpha/KC/CINC-1 IL-3

CXCL10/IP-10/CRG-2 IL-33

CXCL12/SDF-1 alpha IL-4

CXCL13/BLC/BCA-1 IL-5

CXCL16 IL-6

CXCL2/GRO beta/MIP-2/CINC-3 IL-6 R alpha

Cystatin C IL-7

Dkk-1 LDL R

DPPIV/CD26 Leptin/OB

EGF LIX

EGFR M-CSF

EMMPRIN/CD147 MMP-12

Endoglin/CD105 MMP-2

FABP4/A-FABP MMP-3

Complement Factor D/Adipsin MMP-8

13Learn more | rndsystems.com/luminex

Luminex® Assay Customization Tool Build your custom plex from over 450 available analytes — our Luminex Assay Customization Tool makes it simple!

• Design your panel

• Choose a fixed panel

• Select by species

• Share your configuration

• Re-order past panels

• Add to cart

• Purchase

How to Build Your Assay

This video will walk you through the steps necessary to configure and create your Luminex custom assay.

Rat Luminex Assay, Analyte Menu. Cat. # LXSARMHighlighted analytes are unique to R&D Systems

Luminex® AssayCustomization Tool

How to Build Your Assay

Fas Ligand/TNFSF6 MMP-9

FGF basic Nephrin

FGF-21 Oncostatin M/OSM

G-CSF Osteopontin/OPN

Osteoprotegerin/TNFRSF11B Podocalyxin

Pancreatic Polypeptide/PP Prolactin

Proprotein Convertase 9/PCSK9 P-Selectin/CD62P

PDGF-AA RAGE/AGER

PDGF-AB Renin

PDGF-BB Resistin

Periostin/OSF-2 S100A8

PlGF-2 S100A9

Serpin E1/PAI-1 TNF RII/TNFRSF1B

SP-D TNF-alpha

Syndecan-1/CD138 TRANCE/TNFSF11/RANK L

Thrombospondin-4 TWEAK/TNFSF12

TIM-1/KIM-1/HAVCR uPAR

TIMP-1 VEGF

TIMP-4 VEGF R2/KDR/

TNF RI/TNFRSF1A

CXCL2/GRO beta/MIP-2/CINC-3 IL-18/IL-1F4

CXCL3/GRO gamma/CINC-2/DCIP-1 IL-2

GM-CSF IL-4

ICAM-1/CD54 IL-6

IFN-gamma IL-1 alpha/IL-1F1

IL-1 beta/IL-1F2 TIMP-1

IL-10 TNF-alpha

IL-13 VEGF

L-Selectin/CD62L

14 Learn more | rndsystems.com/luminex

Luminex FLEXMAP 3D Magnetic Plate Separator

Luminex® Custom ServicesAccelerate biomarker discovery, validation, and detection with unique analyte panels.

Available Services

• Unique analyte development

• Optimized panel configurations

• Sample type validation

• Evaluation of externally sourced antibodies

• Panel assembly to minimize required sample volume

Need a customized panel. Contact us now.

Accessory Reagents

Luminex Instruments Accesories

Luminex MAGPIX Luminex 100/200

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Which Immunoassay is Right for You?

Description Format Quantitative Sample volume Number of analytes

Proteome Profi ler™ Arrays Membrane Relative 200–500 μL 15–119

Luminex® Assays Bead 25–50 μL Up to 50

Simple Western™ Capillary Cartridge 3 μL >1

DuoSet® ELISAs Flexible 100 μL 1

Quantikine® ELISAs 96-well plate 10–200 μL 1

Simple Plex™ Assays Cartridge 2.5–25 μL Up to 4

B

HRP

BR_Quarterly-Luminex-Update_ASDFY20-8632

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