How to Run an R&D Systems Luminex® Assay

Researchers can encounter a variety of problems when running a Luminex Assay. Explore this page to understand the potential source of your issue and the steps we recommend to correct the problem and run a successful Luminex Assay.

Luminex Assay Protocol

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This video is a general step-by-step guide to running an R&D Systems Luminex Assay. R&D Systems Luminex Assay employ magnetic beads coated with capture antibodies, Phycoerythrin (PE)-conjugated streptavidin is utilized as it binds to the biotinylated detection antibodies. More information is available here (Luminex Assay Principle). Plasma, serum and cell culture supernatant samples are validated for this assay and we recommend that all samples are assayed immediately after they are collected, but samples can be frozen for future use. This kit is for research use only and not for use in diagnostic procedures.

1. Prepare all reagents as instructed.
2. Add 50 µL of standard or sample* to each well.
3. Add 50 µL of diluted Microparticle Cocktail to each well. Incubate for 2 hours at RT on a shaker at 800 rpm.
4. Wash by removing the liquid from each well, filling with 100 µL Wash Buffer, and removing the liquid again. Perform the wash 3 times.
5. Add 50 µL of diluted Biotin- Antibody Cocktail to each well. Cover and incubate for 1 hour at RT on the shaker at 800 rpm.
6. Repeat wash as in step 4.
7. Add 50 µL of diluted Steptavidin-PE to each well. Incubate for 30 minutes at RT on the shaker at 800 rpm.
8. Repeat the wash as in step 4.
9. Add 100 µL of Wash Buffer to each well. Incubate for 2 minutes at RT on the shaker at 800 rpm.
10. Read within 90 minutes using a Luminex or Bio-Rad analyzer. Note: Resuspend microparticles immediately prior to reading.

*Samples may require dilution. See Sample Preparation section.

This concludes our video guide for running an R&D Systems Luminex Assay. For more helpful protocols subscribe to our YouTube channel. Find more information about Luminex Assays and High Performance Assays or contact us.

Luminex Resources

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If you have any questions about your assay please contact R&D Systems Technical Service at techsupport@bio-techne.com.

 Luminex^® is a trademark of Luminex Corporation.