Multiplexing is a revolutionary technique that simultaneously examines and quantifies multiple biological analytes in single sample volumes. Today multiplexing has several applications and is used widely in biomedical and pharmaceutical research.
What are Luminex multiplex bead assays?
Luminex multiplex assay are bead-based immunoassays that employ similar principles to traditional sandwich ELISA assays. In Luminex assays, microspheres are internally dyed with different red and infrared fluorophores. Each of these dyed proportions corresponds to specific bead regions. Luminex multiplex assays help scientists quantify upto 100 analytes in lesser assay volumes than conventional ELISA assays. Scientists employ Luminex multiplex cytokine assays to quantify several biomarkers and cytokines in complex study samples. These assays are critical in providing information regarding diseases and biological processes.
Luminex multiplex assay principle
Analyte-specific antibodies are attached to specific bead regions and then incubated with the study sample. After the initial washing step, the assay sample is incubated with a mix of streptavidin-phycoerythrin reporter molecules and biotinylated detection antibodies. The Luminex system excites the microbeads with one laser and identifies the corresponding bead region. Another laser measures the magnitude of the generated signal. Generally, the PE-derived signal is proportional to the quantity of the bound analyte. Researchers take multiple readings at each bead region to ensure a robust detection.
Benefits of Luminex multiplex assays
Individual assay components are crucial for a robust Luminex multiplex assay. Luminex often employs high-quality antibodies and is tested for multiple clinical and non-clinical applications. Following are some advantages of using Luminex assays.
- Luminex assays can quantify upto 50 analytes in just 25-50 µl of sample volumes. Thus, they save time and resources
- Luminex assays are highly flexible and can be tailored to meet every research need
- Luminex assays are validated similar to gold standard assays for unparalleled precision, accuracy, and sensitivity
- Most importantly, they can analyze numerous unique analytes in complex matrices.
Custom Luminex assays
Similar to traditional immunoassays, Luminex assays provide customizable options for assessing novel analytes. Several Luminex assay services deploy custom assay assistance to develop tailored Luminex assay protocols. Luminex assay services often have a plethora of assays developed against common analytes. However, for novel analytes, they can create a Luminex assay by turning a research idea into a multiplexing solution.
Best practices for Luminex multiplex bead assays
- Bring all reagents to room temperature before beginning assay protocol, unless reagents are required to be kept at cold
- Control and standard samples should never be aliquoted as they are generally single-use
- Whenever possible, use multichannel pipettes for more consistent results. Besides, dispense the study sample by holding the pipette at an angle and avoiding touching the bottom of the well
- Although researchers can use the same pipette tip between replicates, new tips should be used between standards or different samples
- Ideally, researchers must use an automatic washer for a consistent washing step. However, irrespective of using a plate washer or manual washing, keep a 30-second soak interval between washes
- Thoroughly follow the incubation period for each assay step. Do not exceed +/- 5 minutes for every hour of incubation time.
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