Luminex

Luminex Applications: Multiplex Luminex technology allows for simultaneous analysis of a broad selection of analytes (e.g. hormones, cytokines, and adipokines) across a wide range of disease states, including metabolic disease, inflammation, neurodegenerative disease, toxicity, cancer and more.

These multiplexed assays use x-map technology via the MagPix system. The reactants in these assays are attached to the surfaces of tiny, fluorescent, microspheres, each of which carries a unique biological reagent distinguishable by internal dye ratios. Identification of an analyte is based upon specific fluorescent emission spectra of the bead associated with the analyte. Two laser beams with high-speed digital signal processors and computer algorithms distinguish which assay is being carried on each microsphere while quantifying the reaction based on fluorescent reporter signals. This instrumentation allows for the analysis of multiple analytes from a single aliquot of sample.

Multiplex kits are available from several vendors, including Millipore, BioRad, Life Sciences, ThermoFisher, and R&D Systems. Millipore has the most extensive selection but our core has used kits from other companies (the procedures are quite similar despite the vender). See below for the kits that we have the most experience with. Others may be available upon request.

***This core is supported by NIH grants DK059637 (MMPC) and DK020593 (DRTC). Please include an acknowledgment in your publications.***

We have experience with the following assays; others may be available upon request:

Mouse Single Plex Adiponectin - 7 µl - Adiponectin (total)

Mouse Cytokine/Chemokine Panel 1 - 5-60 µl, depending on dilution - Eotaxin/CCL11, G-CSF, GM-CSF, IFN-y, IL-1a, IL-1b, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10/CXCL10, KC/CXCL1, LIF, LIX, MCP-1/CCL2, M-CSF, MIG/CXCL9, MIP-1a/CCL3, MIP-1b/CCL4, MIP-2/CCL8, RANTES/CCL5, TNFa, VEGF

Mouse Pituitary - 25 µl - ACTH, BDNF, FSH, GH, LH, Prolactin, TSH

Mouse/Rat IGF-1 - 10 µl - IGF-1

Mouse Th17 - 60 µl - GM-CSF, INFy, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12(p70), IL-13, IL-15, IL-17A, IL-17E/IL-25, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-28A, IL-28B, IL-31, IL-33, MIP-3a/CCL20, TNFa, TNFb/LTA

We have experience with the following assays; others may be available upon request:

Human Adipokine Panel 1 - 20 µl - Adiponectin (total), Resistin, PAI-1 (total), Lipocalin-2/NGAL, Adipsin

Human Cytokine/Chemokine Panel 1 - 60 µl - EGF, Eotaxin/CCL11, FGF-2, Flt-3 Ligand, Fractalkine/CX3CL1, G-CSF, GM-CSF, GRO, IFNa2, IFN-y, IL-1a, IL-1b, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8CXCL8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1/CCL2/CXDL10, MCP-3/CCL7, MDC/CCL22, MIP-1a/CCL3, MIP-1b/CCL4, PDGF-AA, PDGF-AB/BB, RANTES/CCL5, sCD40L, TGFa, TNFa, TNFb/LTA, VEGF

Human Metabolic Hormone - 60 µl - Active Amylin*, Total Amylin*, C-Peptide, Ghrelin (active)*, GIP (total), GLP-1(active)*, GLP-1 (total)*, Glucagon*, IL-6, Insulin, Leptin, MCP-1/CCL2, Pancreatic Polypeptide, PYY (total), TNFa - *Needs preservative

Human IGF - 60 µl - IGF-1, IGF-2

Human Th17 - 60 µl - GM-CSF, INFy, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12(p70), IL-13, IL-15, IL-17A, IL-17E/IL-25, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-28A, IL-31, IL-33, MIP-3a/CCL20, TNFa, TNFb/LTA

There are general advantages and disadvantages of Luminex assays compared to traditional radioimmunoassays (RIAs) offered by the core.

RIA advantages:

  • Sensitivity
  • Accuracy
  • Cost is typically lower
  • High throughput

RIA disadvantages:

  • More sample volume needed than multiplex
  • Only one analyte may be measured at a time

Luminex advantages:

  • •Small sample volume is especially beneficial for mouse research
  • Ability to analyze up to 100 analytes in a single assay
  • •Customizable (one, all, or any combination of analytes may be chosen within a given panel)
  • •Greater selection of analytes

​Luminex disadvantages:

  • •Not as sensitive as RIA
  • •Not as accurate as RIA
  • •Cost is typically higher (although there is a significant reduction in price for additional analytes run within a single panel)

1.  Who can I contact about  Luminex specific questions?

2.  How does the pricing work? 

  • For Vanderbilt users the first analyte within a panel is $35.08 and each additional analyte within the same panel is $6.00. (Current pricing, including for external users, is available here and in iLab where orders are placed.)
  • For example, to measure three analytes from panel 1 and two analytes from panel 2 the price will be $35.05+6+6 for panel 1 and $35.05+6 for panel 2.

***On submittals for rarely requested panels we require that a minimum number of samples be submitted to cover our costs (e.g. 20). Contact Suzan Vaughan for more information***

3. Can I get a reduced price if I submit more samples? 

  • No. We are an NIH funded non-profit core and we run assays at cost.

4. Do I need to provide the required sample volume for each analyte in the panel? 

  • No. The required sample volume is enough to measure all analytes in the panel simultaneously.

5. What is the required sample volume?

  • See above (this is specific to each assay, typically ranging from 5-60 µl plasma).

6. How long until I get my data? 

  • We process samples in the order that we receive them so it depends on the current demand for the assay. The wait time is typically 2 to 3 weeks once we have the kit and reagents.

***Because Luminex is a custom assay it often requires extra time to order reagents. If you are in a rush, place your order a few weeks before you submit your samples to expedite the process.***

7. What is the sensitivity, cross reactivity, etc of the assay?

  • Vendors supply those details for their individual panels. Please note that analyte concentrations may be below the limits of detection of the assay -- this is out of our control and important to verify ahead of time. 

8. What are the advantages and disadvantages of Luminex assays? 

  • See above.

9. Can I purchase and run my own Luminex panel? 

  • Yes, for a fee Vanderbilt users may use the core's Luminex instrument.