[Skip to Content](https://www.sigmaaldrich.com#main-content) [](https://www.sigmaaldrich.com/CA/en) Products Cart0 CAEN Products [Login / Register](https://www.sigmaaldrich.com/oidc-sign-in) [Order Lookup](https://www.sigmaaldrich.com/CA/en/order-lookup) [Quick Order](https://www.sigmaaldrich.com/CA/en/quick-order) Cart0 [Home](https://www.sigmaaldrich.com/CA/en)[MILLIPLEX® Multiplex Assays for Luminex® Instruments](https://www.sigmaaldrich.com/CA/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/CA/en/technical-documents/product-supporting/milliplex/tips-and-tricks-decide-which-multiplex-assay-fits-your-research)[](https://www.sigmaaldrich.com/CA/en/technical-documents/product-supporting/milliplex/tips-and-tricks-decide-which-multiplex-assay-fits-your-research) - [](https://www.sigmaaldrich.com/CA/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/CA/en/technical-documents/product-supporting/milliplex/luminex-multiplexing-platforms-comparison) - [Species cross-reactivity of MILLIPLEX® multiplex assays](https://www.sigmaaldrich.com/CA/en/technical-documents/product-supporting/milliplex/tips-tricks-species-cross-reactivity) ### Preparation Tips - [Sample preparation techniques](https://www.sigmaaldrich.com/CA/en/technical-documents/product-supporting/milliplex/tips-and-tricks-sample-handling-and-preparation) - [Preparation of reagents](https://www.sigmaaldrich.com/CA/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/CA/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/CA/en/technical-documents/product-supporting/milliplex/tips-tricks-immunoassay-protocol-troubleshooting-advice) - [Equipment settings and data analysis](https://www.sigmaaldrich.com/CA/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/CA/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/CA/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/CA/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/CA/en/technical-documents/product-supporting/milliplex/tips-tricks-species-cross-reactivity) - [Tips & Tricks | Sample Handling and Preparation | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CA/en/technical-documents/product-supporting/milliplex/tips-and-tricks-sample-handling-and-preparation) - [Tips & Tricks | Preparation of Reagents | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CA/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/CA/en/technical-documents/product-supporting/milliplex/tips-tricks-alternate-sample-types-in-cytokine-multiplex-panels) - [Luminex® Multiplexing Platforms Comparison](https://www.sigmaaldrich.com/CA/en/technical-documents/product-supporting/milliplex/luminex-multiplexing-platforms-comparison) - [Comparing xMAP® Sheath Fluid PLUS to Sheath Fluid | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CA/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/CA/en/technical-documents/product-supporting/milliplex/equipment-settings-data-analysis-milliplex-multiplex-assays) - [Tips & Tricks | Immunoassay Protocols & Troubleshooting | MILLIPLEX® Assays](https://www.sigmaaldrich.com/CA/en/technical-documents/product-supporting/milliplex/tips-tricks-immunoassay-protocol-troubleshooting-advice) - [View More](https://www.sigmaaldrich.com/CA/en/search/facet-search?focus=sitecontent&term=facet-search) Top __Sign In To Continue__ To continue reading please sign in or create an account. 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