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[Home](https://www.sigmaaldrich.com/CZ/en)[MILLIPLEX® Multiplex Assays for Luminex® Instruments](https://www.sigmaaldrich.com/CZ/en/products/protein-biology/immunoassay-platform-solutions/milliplex-multiplex-assays-for-luminex-technology)Tips & Tricks Guide to MILLIPLEX® Multiplexing
# Tips & Tricks Guide to MILLIPLEX® Multiplexing
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.
__Section Overview__
- [Industry Guidance for Immunoassays](https://www.sigmaaldrich.com#industryguidance)
- [Multiplexing Tips and Tricks](https://www.sigmaaldrich.com#multiplexingtips)
- [One-Day MILLIPLEX® Multiplex Assays for Limited Lab Time](https://www.sigmaaldrich.com#onedaymilliplex)
- [Request Information](https://www.sigmaaldrich.com#requestinformation)
## [](https://www.sigmaaldrich.com)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](https://www.sigmaaldrich.com#references) below.
## [](https://www.sigmaaldrich.com)Multiplexing Tips and Tricks
Check out these articles for tips and tricks on multiplexing with MILLIPLEX® assays.
### Choosing the Right Assay, Platform, and Materials
- [Decide which MILLIPLEX® assay is right for your research and the materials required](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-and-tricks-decide-which-multiplex-assay-fits-your-research)[](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-and-tricks-decide-which-multiplex-assay-fits-your-research)
- [](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-and-tricks-decide-which-multiplex-assay-fits-your-research)[Choose the best Luminex® multiplex platform for your research](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/luminex-multiplexing-platforms-comparison)
- [Species cross-reactivity of MILLIPLEX® multiplex assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-species-cross-reactivity)
### Preparation Tips
- [Sample preparation techniques](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-and-tricks-sample-handling-and-preparation)
- [Preparation of reagents](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-preparation-of-reagents-milliplex-multiplex-assays)
- [Using MILLIPLEX® kits in alternate sample types](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-alternate-sample-types-in-cytokine-multiplex-panels)
### Running Assays and Analyzing Data
- [Immunoassay procedures and troubleshooting tips](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-immunoassay-protocol-troubleshooting-advice)
- [Equipment settings and data analysis](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/equipment-settings-data-analysis-milliplex-multiplex-assays)
### General Questions on MILLIPLEX® Assays
- [MILLIPLEX® Multiplex Assays FAQs](https://www.sigmaaldrich.com/CZ/en/technical-documents/technical-article/protein-biology/immunohistochemistry/milliplex-faq)
## [](https://www.sigmaaldrich.com)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 required[4](https://www.sigmaaldrich.com#references), 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](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/multiplexing-in-vet-med-and-animal-health-research). Select single-day assays are included below.
## Immunology/Inflammation Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## Virology Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## Metabolism/Endocrinology Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## Cardiovascular Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## Bone Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## Neuroscience Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## Toxicity Assays
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
__For Research Use Only. Not For Use In Diagnostic Procedures.__
## Request Information
Have more questions about MILLIPLEX® multiplex assays? Fill out the form by clicking on the button below.
[Request info on MILLIPLEX® assays](https://www.sigmaaldrich.com/CZ/en/campaigns/milliplex-request-info)
### 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](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](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](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](https://doi.org/10.3389/fimmu.2020.572634)
__Related Articles__
- [Tips & Tricks | Species Cross-Reactivity | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-species-cross-reactivity)
- [Tips & Tricks | Sample Handling and Preparation | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-and-tricks-sample-handling-and-preparation)
- [Tips & Tricks | Preparation of Reagents | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-preparation-of-reagents-milliplex-multiplex-assays)
- [Tips & Tricks | Alternate Sample Types in MILLIPLEX® Cytokine Multiplex Panels](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-alternate-sample-types-in-cytokine-multiplex-panels)
- [Luminex® Multiplexing Platforms Comparison](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/luminex-multiplexing-platforms-comparison)
- [Comparing xMAP® Sheath Fluid PLUS to Sheath Fluid | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/comparing-xmap-sheath-fluid-plus-to-sheath-fluid)
- [Tips & Tricks | Equipment Settings & Data Analysis | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/equipment-settings-data-analysis-milliplex-multiplex-assays)
- [Tips & Tricks | Immunoassay Protocols & Troubleshooting | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CZ/en/technical-documents/product-supporting/milliplex/tips-tricks-immunoassay-protocol-troubleshooting-advice)
- [View More](https://www.sigmaaldrich.com/CZ/en/search/facet-search?focus=sitecontent&term=facet-search)
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