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Multiplexing Tips & Tricks

We’re so confident in the benefits of MILLIPLEX® multiplex kits that we’ve compiled tips and tricks, straight from the experts, to eliminate any doubt in your ability to multiplex like a pro. Enhance the power of your research with MILLIPLEX® multiplexing by learning more about industry guidance, multiplexing tips, one-day multiplex assays for quick turnaround times, and more.

Industry Guidance for Immunoassays

Let industry guidance lead you to MILLIPLEX® multiplexing. From academia to contract research to pharma, MILLIPLEX® kits meet the ever-increasing demand for high-quality immunoassays for reproducible results. This includes:

  • Detection and sensitivity
  • Performance in a sample matrix
  • Specificity
  • Selectivity
  • Precision and accuracy
  • Linearity
  • Stability
  • Cross-talk
  • Lot-to-lot reproducibility
  • Vendor support

Want to learn more about industry guidance on assay development and validation? We recommend these references1,2,3 below.

One-Day MILLIPLEX® Multiplex Assays for Limited Lab Time

Many of our MILLIPLEX® multiplex assays can be completed in a single day by using a shorter primary incubation time. With only 90 minutes of hands-on time required4, these assays allow for more time to work on other important laboratory projects. Results are available in 4-5 hours, depending on incubation times, making them ideal for limited laboratory time.

The typical MILLIPLEX® one-day assay workflow is as follows:

  • Step 1: Set up plate with samples/standards, beads, buffer/matrix. Incubate at room temperature for 1-2 hours*.
  • Step 2: Wash plate. Add detection antibodies. Incubate at room temperature for 1 hour*.
  • Step 3: Wash plate*. Add Streptavidin-Phycoerythrin. Incubate at room temperature for 30 minutes.
  • Step 4: Wash plate. Read on Luminex® instrument.

*See protocol for specific incubation times and wash steps.

More than 50 MILLIPLEX® multiplex kits, covering several research areas, are available with a single-day protocol. This includes more than 30 immunology assays, as well as virology, metabolism/endocrinology, cardiovascular, neuroscience, and more. MILLIPLEX® multiplex assays spanning multiple species can be completed in one day, including human, mouse, rat, non-human primate, and many of our veterinary medicine and animal health panels. Select single-day assays are included below.

Immunology/Inflammation Assays
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Virology Assays
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Metabolism/Endocrinology Assays
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Cardiovascular Assays
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Bone Assays
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Neuroscience Assays
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Toxicity Assays
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For Research Use Only. Not For Use In Diagnostic Procedures.

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References

1.
Lee JW, Devanarayan V, Barrett YC, Weiner R, Allinson J, Fountain S, Keller S, Weinryb I, Green M, Duan L, et al. 2006. Fit-for-Purpose Method Development and Validation for Successful Biomarker Measurement. Pharm Res. 23(2):312-328. https://doi.org/10.1007/s11095-005-9045-3
2.
Jani D, Allinson J, Berisha F, Cowan KJ, Devanarayan V, Gleason C, Jeromin A, Keller S, Khan MU, Nowatzke B, et al. 2016. Recommendations for Use and Fit-for-Purpose Validation of Biomarker Multiplex Ligand Binding Assays in Drug Development. AAPS J. 18(1):1-14. https://doi.org/10.1208/s12248-015-9820-y
3.
Andreasson U, Perret-Liaudet A, van Waalwijk van Doorn LJC, Blennow K, Chiasserini D, Engelborghs S, Fladby T, Genc S, Kruse N, Kuiperij HB, et al. A Practical Guide to Immunoassay Method Validation. Front. Neurol.. 6 https://doi.org/10.3389/fneur.2015.00179
4.
Günther A, Becker M, Göpfert J, Joos T, Schneiderhan-Marra N. Comparison of Bead-Based Fluorescence Versus Planar Electrochemiluminescence Multiplex Immunoassays for Measuring Cytokines in Human Plasma. Front. Immunol.. 11 https://doi.org/10.3389/fimmu.2020.572634
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