MILLIPLEX^® Multiplex Assays for Luminex^® Instruments FAQs

Have questions about MILLIPLEX^® multiplex assays for Luminex^® instruments? Find the answers to frequently asked questions below straight from our team of experts.

Do you have a resource guide for using MILLIPLEX^® assay panels?

For the most current and comprehensive catalog of MILLIPLEX^® products, download a copy of our Analyte Quarterly to see our entire portfolio of MILLIPLEX^® assays, Luminex^® instruments, and all of our immunoassay solutions and offerings, from ultrasensitive detection with Single Molecule Counting (SMC^®) technology to our robust selection of ELISA kits.

Additionally, our Tips & Tricks provides technical information on MILLIPLEX^® assays and Luminex^® instruments, including information on the technology and how it works, protocols, data analysis, and species cross-reactivity data for our most popular analytes.

What is included in every quantitative MILLIPLEX^® multiplex kit?

Figure 1 shows all the components included in every quantitative MILLIPLEX^® multiplex kit.

Figure 1 All components are included in every quantitative MILLIPLEX^® multiplex kit, including: (A) 10X Wash Buffer, (B) Mixing Bottle, (C) Magnetic Beads—Capture antibody, conjugated beads in solution, (D) Standard, (E) Controls—QC1 & QC2, (F) Assay Buffer, (G) Bead Diluent, (H) Serum Matrix, (I) SAPE—Streptavidin Phycoerythrin, (J) Plate Sealers, (K) 96-Well Plate—Greiner Black Mylar, and (L) Detection Antibody Cocktail.

This includes:

• Luminex^® Magnetic Beads
  • Antibody-conjugated beads in solution (capture or primary antibody)
  • Premixed formats available for select kits
• Bead Diluent
  • For mixing the beads to the correct volume
  • Contains a cocktail of proprietary blocking reagents which significantly reduces false signal to ensure true biomarker measurement (including that from heterophilic antibodies)
• Standards
  • Lyophilized cocktail of recombinant protein analytes
  • Standard dilution curve is used to calibrate the assay
• Quality Controls (QCs)
  • High and low QCs and Range Sheet are provided to ensure that proper pipetting and assay setup has been accomplished
  • Customers should include appropriate Sample QCs in each assay
• Serum Matrix Solution
  • Lyophilized serum matrix provided for dilution of standards when using serum and plasma samples
  • Provides a more accurate analyte quantification than a standard curve diluted in buffer; Serum matrix simulates the conditions in which the native protein is analyzed
• Detection Antibody Cocktail
  • Contains multiple biotinylated reporter (detection) antibodies in solution ready to use
• SA-PE
  • Streptavidin Phycoerythrin (a fluorophore) comes ready to use
• Assay Buffer
• Wash Buffers
• Plates and Sealers
• Protocol

Included with Cell Signaling Assays:

• Lysis Buffer
• Cell Lysates for use as positive and negative controls in select kits

Included with Antibody Detection Assays

• PE-Ig conjugate
• Qualitative assays do not include standard and QCs

How many analytes and detection kits do you offer?

We offer over 500 unique circulating analytes, over 100 intracellular analytes, and more than 130 kits with more than 35 kits premixed for your convenience. Our kits also cover the broadest range of species: human, non-human primate, mouse, rat, canine, feline, porcine, ovine, bovine, chicken, and equine.

For a full list of our current offerings, download our Analyte Quarterly brochure.

Who do I contact if I need additional assistance about an order, product price and availability, or login assistance?

Please contact our Customer Service.

Who do I contact if I have technical or scientific questions or concerns about a MILLIPLEX^® assay?

Please follow the link to contact Technical Support by phone or send a message to Technical Support.

Who do I contact if I need to request a call or visit from my Biomarker Sales Specialist?

To request a call or visit from your Biomarker Sales Specialist please send a message to the Sales Team.

Who can I contact if I need to replace a reagent in a MILLIPLEX^® kit?

Please contact our Technical Support or your Sales Representative.

Where can I find more information if I am planning a large, long-term study or writing a grant proposal for funding?

Request a Sales Specialist to contact you to assist with your immunoassay needs, including assay validation requirements, custom assays, supply chain planning, quality agreements, packaging and shipping, and reagent change notifications.

Where can I find more information about your custom assays for new analytes or to combine analytes for multiple MILLIPLEX^® kits?

For a custom MILLIPLEX^® multiplex assay or ELISA, please visit our Custom Assays page and fill out the form.

What kind of quality and assay verification do you provide with your MILLIPLEX^® kits?

MILLIPLEX^® kits are manufactured in facilities that are ISO 9001-2015 compliant and are for research use only (RUO), and not for use in clinical or for medical diagnostic purposes. Discover how our quality differentiates our MILLIPLEX^® assays from development to manufacturing.

MILLIPLEX^® kits are tested for assay sensitivity, cross-reactivity, precision, and more. Table 1 describes the criteria.

Table 1. Criteria tested for MILLIPLEX® multiplex kits.

Criteria	Description
Verified Sample Matrix	Serum, plasma, or tissue culture supernatants (or other sample types as indicated in the kit protocol)
Assay Sensitivity	Within biological significance; minDC reported
Selectivity/Specificity	Cross-reactivity between antibodies is non-detectable or negligible, and antibodies are verified
Precision	Intra-assay within 15% CV (SD/mean)   Inter-assay within 20% CV (SD/mean)
Accuracy	Within ± 30%
Linearity	Within ± 30%
Kit Shipping Stability	At 37 °C, 4 °C, and Room Temperature for 7 Days
Method Comparison	When available, against in-house or other vendors
Species Cross-Reactivity	Tested against available samples (contact Technical Support or visit our Species Cross-Reactivity page for information)

Are your MILLIPLEX^® kits and analyte multiplexing kits customizable?

Most of our MILLIPLEX^® kits are customizable, often referred to as “configurable,” which means you may choose one, any number, or all of the analytes listed for your research, and you only pay for the analytes you select.

Can I mix reagents from more than one of the same type of kit?

Please only mix reagents if the kit reagents are all from the same lot (lot numbers are on each bottle label inside the kit) and the reagents have been stored under the recommended conditions.

Can two or more MILLIPLEX^® kits be combined?

Each kit is developed and verified to optimally perform with the included reagents and according to protocol recommendations. Combining kits can risk antibody cross-reactivity, incompatible sample dilution factors, inappropriate serum matrix, buffers, or incubation periods – any of which can skew your results.

If you wish to combine analytes from different kits, check out our Custom Assays. You can rely on our expertise to help you find the best options for your research needs.

Can I use a kit meant for one species to detect samples from another species?

Our Species Cross-Reactivity article contains analyte species cross-reactivity data for select kits. You may also contact our Technical Support team.

Can a MILLIPLEX^® kit be used to detect an analyte in saliva, urine, or tears?

Our kits have been tested in the sample types indicated in each kit protocol. Typically, our kits are tested for use with plasma or serum samples, cell culture samples, or lysate samples.

Some of our kits have been specifically tested for urine or CSF (cerebral spinal fluid). Before trying our kits with a new sample type, please contact Technical Support for recommendations or our Custom Assays team can run verification testing on your specific sample type.

For tips on sample prep for MILLIPLEX^® assays , check out our Sample Preparation and Handling article.

How does the price of multiplexing compare to running ELISAs?

Each MILLIPLEX^® multiplex kit has a base price for the kit with one analyte. Each additional analyte adds incrementally to the price. If you only wish to detect one or two analytes, two separate single analyte ELISA kits may suit your budget and research needs. However, when you interrogate multiple analytes, multiplexing becomes a real cost-saver.

If your goal is to save precious sample volume, then multiplexing is key. Most of our MILLIPLEX^® kits require only 25 µL of sample, sometimes “neat” and sometimes diluted, according to each protocol. You save time and sample by running just one easy assay to quantify all your selected analytes of interest. Our kits come with all the reagents you need to complete your assay, including the assay plate and sealers. Just add water, sample, and follow the protocol.

Can I use any plate reader for MILLIPLEX^® assays?

A Luminex^® instrument is required to run Luminex^® technology-based immunoassays, including MILLIPLEX^® assays and kits. There are several instruments currently available for use with MILLIPLEX^® kits:

• xMAP^® INTELLIFLEX^® System RUO
• xMAP^® INTELLIFLEX^® System DRSE
• FLEXMAP 3D^® System
• Luminex^® 200™ System

For more information on the differences between these instruments, visit our Luminex^® Multiplexing Platforms Comparison page. These instruments may be purchased from us or other Luminex^® partners. The Bio-Plex^® instrument may also be used to run any Luminex^® technology-based immunoassay including MILLIPLEX^® assays.

How do I cite MILLIPLEX^® multiplex assays in my publications?

• If you are based in the United States or Canada, state MilliporeSigma as the source of your assay.
• If you are based anywhere in the world outside of the United States or Canada, state Merck KGaA, Darmstadt, Germany, as the source of your assay.
• Include the full MILLIPLEX^® kit name and kit catalog number, and also list the analytes you used from the kit.
• Be sure to include the species of samples you used and how you diluted your samples.

For Research Use Only. Not For Use In Diagnostic Procedures.