MILLIPLEX® Multiplex Assays Using Luminex®

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Tips and TricksThe Power of Multiplex Biomarker Analysis from the makers of MILLIPLEX® MAP

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Featured Customer Story
Merck:/Freestyle/BI-Bioscience/Protein-Detection/bmia-images/Mukesh-150px.jpgInterested in how MILLIPLEX® MAP assays can further Zika research?

The MILLIPLEX® MAP Rat Cytokine Panel (Cat. No. RECYTMAG-65K) allowed Dr. Mukesh Kumar, of the University of Hawaii, to develop a guinea pig model for Zika virus infection. Learn more about his research by viewing the video below, and read his most recent publication.

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Characterizing the maternal immune environment during pregnancy: Implications for autism spectrum disorders

Speakers: Dr. Karen L. Jones, University of California-Davis, and Dr. Anthony Saporita, Merck R&D

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Dr. Karen L. Jones Dr. Anthony Saporita

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Looking for a little more information on autism and the immune system?

Read the SelectScience article about Dr. Karen L. Jones’ research:
Autism Linked to Abnormal Immune Responses of Pregnant Mothers: Learn how the maternal immune profile of mothers can predict neurodevelopmental abnormalities

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Need Tips on Optimizing your Multiplex Assays?
Merck:/Freestyle/BI-Bioscience/Protein-Detection/bmia-images/Sonali-Nayak-headshot.jpgRead the SelectScience Interview with Sonali Nayak, Bioscience Technical Services Manager, Merck:
Discover How Multiplex Assays are Advancing Biomedical Research: Top tips to optimize your analyses 

Get Sonali Nayak’s take on how to best run your MILLIPLEX® MAP assays.

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As your research uncovers the significant role of more and more biomarkers in both normal and disease states, you need a reliable partner for quality assays and consistent performance. We not only offer you the largest portfolio of biomarker assays—including ELISAs, RIAs and MILLIPLEX® MAP (multi-analyte profiling), our kits and reagents provide you with consistent, high-quality tools so that you can do your best work while saving time, labor and cost.

In addition to the assay specifications listed in the protocol, we evaluate other performance criteria during our validation process: cross-reactivity, dilution linearity, kit stability and sample behavior (e.g. detectability and stability).

Each MILLIPLEX® MAP kit includes:
  • Quality controls (QCs) provided to qualify assay performance
  • Comparison of standard (calibrator) and QC lots to a reference lot to ensure lot-to-lot consistency
  • Optimized serum matrix to mimic native analyte environment
  • Detection antibody cocktails designed to yield consistent analyte profiles within panel

How to Decide Which MILLIPLEX® Assay Is Best for Your Research

In Method Comparison tests (comparing ELISA and MILLIPLEX® MAP  assays), while the absolute values are not exactly the same, the results do correlate. Hence when switching from one assay method to another, a correction factor may be used when comparing with past data. In most situations the dynamic range and the sensitivity of the assay may be better with the MILLIPLEX® kits.

Please contact Technical Support for more information on correlation factors.

To locate protocols and technical documents for a specific panel, choose any of these options:
  • Search the website for the catalog number.

  • The link to the protocol can be found using:
    • The “Documents” tab
    • The right-hand side of the product name and catalog number
    • The top of the Product Description page
  • To find a panel that contains the analytes you want to measure, use the Analyte Kit Finder at the bottom of this page. Alternatively, type the name of the analyte into the search bar at and use the filters at left to refine your search for “Multiplexing.”

How to Find More Technical Information About Each MILLIPLEX® Assay

  • To find publications citing a specific panel or analyte:
    • Search the website for the catalog number. Links to some of the references using the product can be found at the top of the Product Description Page
    • For a more complete list, contact technical support
  • To determine cross-reactivity for other species for a panel or analyte see the Species Cross-reactivity Tables in our technical guide
  • For intracellular assay kits, we analytically validate the assay only with human cell/tissue culture samples. However, we provide the species homology for each analyte in a table on the product detail page on our website. We have also compiled a list of this information on our website:

    Kits - Kit Species Cross Reactivity

    MAPmate™ Species Cross Reactivity Table

Discover the Latest in New Multiplex and Single Plex Protein Detection — New Multiplex Kits!

384-Well Human High Sensitivity T Cell (21 plex) Assay

(Cat. No. HSTC384-28K)
(Cat. No. HSTCMAG384-PX21)
(Cat. No. HSTCMAG384PX21BK)
Introducing kits for your FLEXMAP 3D® instrument: get more with 384
Now you can perform your human cytokine assays faster and more economically with the new MILLIPLEX® MAPHuman High Sensitivity T-Cell 21-plex assay for 384-well plates.
Benefits of 384-well kits:
  • Analytically validated to meet the same QC criteria as our 96-well kits. 
  • You get everything you need — all in one kit — including the confidence in your data that you’ve come to expect from MILLIPLEX® assays. 
  • One standard curve set for each plate
  • Obtain up to 366 sample data points per plate (183 samples run in duplicate) in the same amount of time it would take you to run a 96-well plate
Convert existing kits to 384-well format
Our Custom Assay Development Services can make any existing MILLIPLEX® kit into a 384-well kit within a few weeks, including all species and kits. Just let us know what you need and when you need it, and we will develop a kit that is right for your laboratory.

View data showing comparable sensitivity between 384-well and 96-well assay kits.

Analyte96-well MinDC384-well MinDC
Fractalkine 12.20 6.00
GM-CSF 2.30 0.40
IFNg 0.22 0.17
1L-1b 0.03 0.13
1L-2 0.02 0.09
1L-4 0.01 0.56
1L-5 0.03 0.21
1L-6 0.02 0.04
1L-7 0.94 0.20
1L-8 0.04 0.23
IL12p70 0.03 0.17
IL-10 0.59 0.70
1L-13 0.06 0.06
1L-17A 0.02 0.30
1L-21 0.06 0.04
1L-23 0.26 0.29
ITAC 2.40 0.50
MIP1a 0.10 0.05
MIP1b 3.70 1.70
MIP3 0.52 0.19
TFNa 0.02 0.13
A comparison of MinDCs between the 96-well MILLIPLEX MAP Human High Sensitivity T Cell Premix kit (Cat. No. HSTCMAG28SPMX21) and the 384-well MILLIPLEX MAP Human High Sensitivity T Cell Premix kit (Cat. No. HSTCMAG384-PX21).

Phospho/Total AKT 2-Plexes

The three Akt isoforms (Akt1, Akt2, and Akt3) may have unique, isoform-specific roles in differentiation, proliferation, and other processes. To measure relative levels of phospho- and total Akt1, Akt2, and Akt3, we developed MILLIPLEX® MAP Phospho/Total 2-plex assays for each of the Akt isoforms. Each 2-plex assay enables the measurement of both phosphorylated and total protein in the same well. In addition, each Phospho/Total 2-plex assay can be combined with any other (non-Akt) Phospho/Total 2 plex.

Data obtained using the Phospho/Total Akt Isoform 2-plex Assays are shown below for SH-SY5Y cells. To view MCF7 cell data, read the latest Analyte Quarterly brochure.

Merck:/Freestyle/BI-Bioscience/Protein-Detection/bmia-images/phospo-graph.jpgData obtained using MILLIPLEX® MAP Cell Signaling Phospho/Total Akt isoform 2-plex assays with lysates of stimulated SH-SY5Y cells. MILLIPLEX® MAP Cell Signaling Phospho/Total Akt isoform 2-plex assays were pooled and used to evaluate the cellular response to retinoic acid (RA), which induces neuronal differentiation of SH-SY5Y cells. After 3 days, cells cultured in the presence or absence of RA were collected and lysed according to assay protocols. MFI measurements were obtained, and the ratios of phosphoprotein to total protein were calculated for each, Phospho/Total Akt1, Akt2 and Akt3 2 plexes.

Human Diabetes Premixed Panel

Use this new premixed panel to simultaneously quantify five analytes important in studies of diabetes and obesity: Insulin, Glucagon, Leptin, Glucagon-like peptide (GLP-1), and C-Peptide. This panel is a subset of our popular Human Metabolic Hormone Panel (HMHEMAG-34K) and exhibits high sensitivity, consistency, and reproducibility.

Learn More about Human Diabetes Premixed Panel
Comparison of sensitivity, sample volume requirements, and standard curve range vs. electrochemiluminescence (ECL) detection platform
(ECL) pg/mL
Sample volume
Sample volume
Dynamic Range
Dynamic Range
Glucagon 20 9.6 40 μL 25 μL 2.4-10,000 13.7-10,000
Leptin 22 38.5 24-100,000 137-100,000
GLP-1 (active) 1.0 1.1 2.4-10,000 2.7-2,000
Insulin 9.0 22.1 12.2-50,000 137-100,000
C-Peptide n/a 8.4 n/a 27.4-20,000

Human Liver Injury Panel

This panel is a five-plex assay kit to be used for the simultaneous detection of hepatotoxicity biomarkers, liver-type arginase 1 (ARG1), α-glutathione-S-transferase (GST-α), 5’nucleotidase (5’NT), sorbitol dehydrogenase (SDH), and malate dehydrogenase 1 (MDH1), in serum or plasma samples. These biomarkers are hepatocellular and hepatobiliary enzymes that are released into circulation upon occurrence of liver injury. The sensitivity of this multiplex assay panel is illustrated in the table below.

Learn More about Human Liver Injury Panel

AnalyteMinDC (pg/mL)MinDC+2SD (pg/mL)
ARG1 26.4 70.7
MDH1 16.7 34.9
GSTα 2.6 4.0
5’-NT 15.0 40.4
SDH 107.0 238.4

Human RTK Phosphoprotein Panel

RTKs (Receptor Tyrosine Kinases) are transmembrane proteins that act as receptors for growth factors, neurotrophic factors and other extracellular signaling molecules. RTKs play important, interconnected roles in multiple biological pathways. Unlike traditional cell signaling assays, which are limited to measuring individual RTKs at a time, multiplex immunodetection enables you to screen multiple RTK-mediated signaling pathways at once, so you can quickly determine on which pathway to focus your study.

Learn More about Human RTK Phosphoprotein Panel


Request the Analyte Quarterly brochure for a complete, updated list of assays.

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Multiplex, ELISA and RIA Assay Finder

Biomarkers play a significant role in both normal and disease states and you need a reliable partner for quality assays and consistent performance. Our multiplex, ELISA and RIA biomarker assays provide you with consistent, high-quality tools so that you can do your best work while saving time, labor and cost. Use the assay finder below to choose the correct assay for your biomarker of interest, so you can obtain the best quality data the first time.
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Design & Purchase Your Own Kit

Already know your analytes of choice? Start designing and purchasing your own kits now!

Choose Customizable Panels & Premixed Kits – OR – Cell Signaling MAPmates™ Assays.

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Multiplex Customer Testimonial

What are researchers saying about MILLIPLEX® MAP Quality?

“I have used Merck MILLIPLEX® MAP kits for the past four years. Initially I started using these kits to multiplex a specific set of markers, but given their reliability, consistency, and ease of use, I have transitioned all my experiments away from traditional ELISAs. Beyond just being a great product, Merck's customer and technical support is at the top of the industry. The combination of superior product with award-winning support has extended the capabilities of my research, allowing me to answer new novel research questions. I consider MILLIPLEX® MAP kits an important component of my research toolbox.” 

Brian McFarlin, PhD, FACSM, University of North Texas, Denton, Texas

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