Human Th9/Th17/Th22 Luminex® Performance Assay 18-plex Fixed Panel
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Human Th9/Th17/Th22 Luminex® Performance Assay 18-plex Fixed Panel Summary
| Panel | Luminex Performance Human Fixed Th9/Th17/Th22 Panel |
| Assay Type | Magnetic bead-based multiplex assay for the Luminex® platform |
| Format | 1 x 96-well microplate and Magnetic antibody-coated beads |
| Species | Human |
| Analytes Detected | Please see analyte list in assay customization tool below. |
| Performance Validation | Our Luminex High Performance Assays undergo our most extensive validation testing. All Luminex High Performance Assay Panels are validated for use with serum and plasma samples. Some High Performance Assays are additionally validated for cell culture supernatants, milk, saliva, or urine, as indicated on the individual product data sheets. Luminex High Performance Assays are tested for sensitivity (three-quarters the low standard), intra-assay precision, inter-assay precision, and to ensure assay linearity for validated sample types. Recovery values for individual samples in validated sample types are also tested. Validation data for each analyte can be found in the product datasheet. |
Not finding your analyte of interest? View our complete Luminex offering in the full version of our Luminex Assay Customization Tool. We also offer Custom Luminex Services.
Magnetic Luminex High Performance Assays are flexible bead-based multiplex assays. They allow up to 50 user-defined target analytes to be simultaneously profiled using cell culture supernates, serum, or plasma samples. Some panels are also validated with human milk, saliva, and/or urine samples.
- Simultaneously profile analytes in one sample
- Magnetic format allows for easier wash steps
- Undergoes similar validation testing as Quantikine® ELISA Kits
- Precise and Specific
- Requires a small sample volume (<50 µL)
- Run your assay in just 3-5 hours
- Mass-calibrated standards for consistent results with every new lot of material
- Enough reagents for one 96-well plate
- Premixed cocktail of antibody-coated Magnetic beads
- Premixed cocktail of biotinylated detection antibody
- Standard Cocktail(s)
- Bead Diluent
- Biotin Antibody Diluent
- Calibrator Diluent
- Wash Buffer
- Streptavidin-PE
- One flat-bottom 96-well Microplate
- Foil Plate Sealers (4)
- Mixing Bottle
- Standard Value Card**
**A standard curve must be generated each time an assay is run, utilizing values from the Standard Value Card included in the Kit.
Precision
|
Intra-Assay Precision - is generated from the mean of the %CV's from 40 reportable results across two different concentrations of analytes in a single serum assay Inter-Assay Precision - is generated from the mean of the %CV's across two different concentrations of analytes across 30 different serum assays. |
||
| n=40 (20x2 levels) | n=30 | |
|---|---|---|
| Analyte | Intra-Assay (%CV) | Inter-Assay (%CV) |
| CD40 Ligand | 3.8 | 10.8 |
| GM-CSF | 3.4 | 11.2 |
| IFN-gamma | 2.3 | 10.9 |
| IL-1 beta | 2.8 | 10.1 |
| IL-2 | 3.9 | 9.6 |
| IL-4 | 2.4 | 8.4 |
| IL-5 | 1.8 | 8.3 |
| IL-6 | 2.2 | 8.8 |
| IL-9 | 6.2 | 10.3 |
| IL-10 | 3.6 | 11.2 |
| IL-12 p70 | 2.6 | 9.8 |
| IL-13 | 2.5 | 8.3 |
| IL-15 | 3.2 | 11.2 |
| IL-17A | 2.5 | 9.8 |
| IL-17E | 3.1 | 8.1 |
| IL-33 | 2.9 | 9.7 |
| MIP-3 alpha | 3.0 | 9.4 |
| TNF-alpha | 3.0 | 10.3 |
Accuracy
|
Linearity - The data represents mean spiked or natural linearity in serum matrix samples. Spike Recovery - The data represents mean percent recovery of spiked standards ranging from low, medium, and high concentration in serum matrix samples. |
||||
| Linearity | Spiked Recovery |
|||
|---|---|---|---|---|
| Analyte | N | Average % | N | Average % |
| CD40 Ligand | 4 | 131 | 4 | 75 |
| GM-CSF | 4 | 101 | 4 | 98 |
| IFN-gamma | 4 | 127 | 4 | 73 |
| IL-1 beta | 4 | 106 | 4 | 94 |
| IL-2 | 4 | 115 | 4 | 88 |
| IL-4 | 4 | 141 | 4 | 58 |
| IL-5 | 4 | 113 | 4 | 88 |
| IL-6 | 4 | 126 | 4 | 78 |
| IL-9 | 4 | 111 | 4 | 96 |
| IL-10 | 4 | 117 | 4 | 90 |
| IL-12 p70 | 4 | 111 | 4 | 87 |
| IL-13 | 4 | 118 | 4 | 89 |
| IL-15 | 4 | 103 | 4 | 76 |
| IL-17A | 4 | 128 | 4 | 72 |
| IL-17E | 4 | 15 | 4 | 55 |
| IL-33 | 4 | 103 | 4 | 87 |
| MIP-3 alpha | 4 | 115 | 4 | 99 |
| TNF-alpha | 4 | 134 | 4 | 71 |
Assays for the Luminex platform are offered as High Performance Assays or Assays. The Luminex High Performance Assays are fully validated panels with a focused selection of analytes. They are available in We Mix, You Mix, and Predetermined formats. The Luminex Assays allows for the maximum number of analytes in a multiplex and is supplied as a premixed kit. View a table comparing the features and benefits of these bead-based multiplex assays.
Magnetic Luminex High Performance Assays are designed for use with the Luminex MAGPIX CCD Imager. Alternatively, kits can be used with the Luminex 100/200™ or FLEXMAP 3D, dual laser, flow-based sorting and detection platforms.
Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. One laser is bead-specific and determines which analyte is being detected. A magnet in the analyzer captures and holds the superparaMagnetic microparticles in a monolayer. Two spectrally distinct Light Emitting Diodes (LEDs) illuminate the beads. One LED identifies the analyte that is being detected and the second LED determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of analyte bound. Each well is imaged with a CCD camera. Kits can also be used with Luminex 100/200 or a Bio-Rad Bio-Plex dual laser, flow-based systems.
Scientific Data
Background: Th9/Th17/Th22 Assay Kits
T helper type 17 (Th17) cells are involved in the immune response mounted against specific fungi and extracellular bacteria. In mice, Th17 cells develop from naive CD4+ T cells in the presence of TGF-beta and IL-6. These cytokines induce the STAT3-dependent expression of IL-21, IL-23 R, and the transcription factor, ROR gamma t. IL-21 and IL-23 regulate the establishment and clonal expansion of Th17 cells, while ROR gamma t-induced gene expression leads to the secretion of IL-17A, IL-17F, and IL-22. Cytokines secreted by Th17 cells stimulate chemokine secretion by resident cells, leading to the recruitment of neutrophils and macrophages to sites of inflammation. These cells, in turn, produce additional cytokines and proteases that further exacerbate the immune response. In contrast to mouse Th17 differentiation, Th17 polarization in humans requires IL-1 beta, IL-6, IL-21, and IL-23, but seems to be less dependent upon TGF-beta. One other notable difference is that human Th17 cells secrete IL-26, an IL-10 family cytokine without a murine homologue. Cytokines produced by Th17 cells can have both beneficial and pathogenic effects. While they play a central role in eliminating harmful microbes, persistent secretion of Th17 cytokines promotes chronic inflammation and may be involved in the pathogenesis of inflammatory and autoimmune diseases, including rheumatoid arthritis, multiple sclerosis, and inflammatory bowel disorders.
Product Datasheets
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