Human IL-17E/IL-25 XL Luminex® Performance Assay

Catalog #: LUXLM1258 Datasheet / COA / SDS

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IL-17E/IL-25 XL Cytokine Luminex Panel Standard Curves
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Human IL-17E/IL-25 XL Luminex® Performance Assay Summary

Panel Luminex Performance Human XL Cytokine 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.

View Luminex High Performance Human XL validation data.

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Assays

Below are kits that are available based on your selections. These selections may be further customized before adding to the cart or emailing the configuration to your purchasing department or local distributor to place your order. Selected analytes may be included in more than one High Performance Assay. All assay options are displayed.

If you run into bead region incompatibilities, send your Luminex code to our team and find out more.

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Product Features

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 Human IL-17E/IL-25 with up to analytes of your choice 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
Base Kit Components
  • 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**
  • High and low controls with Control Mean Value Card

**A standard curve must be generated each time an assay is run, utilizing values from the Standard Value Card included in the Base Kit.

Bead Set Contents
  • User-mix antibody-coated Magnetic beads in individual vials

Recommended Sample Types: Cell culture supernates, serum, EDTA plasma, and heparin plasma

Components: Human IL-17E/IL-25 Magnetic Microparticles (Part 898810) is supplied as a 100X concentrated stock (0.075 mL) with preservatives.

Other Supplies Required: Magnetic Luminex® Performance Assay Human XL Discovery Base Kit (R&D Systems®, Catalog # LUXLM000).

Sensitivity: The Minimum Detectable Dose (MDD) was determined by adding two standard deviations to the mean MFI of twenty zero standard replicates and calculating the corresponding concentration.
Six assays were evaluated, and the MDD of human IL-17E ranged from 1.88-4.95 pg/mL. The mean MDD was 1.95 pg/mL.

Specificity: This assay recognizes natural and recombinant human IL-17E.Recombinant human IL-17B R interferes at concentrations > 0.823 ng/mL.

Bead Region: 51
 

Precision

Intra-assay Precision (precision within an assay) - Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (precision between assays) - Two samples of known concentration were tested in thirty separate assays to assess precision between assays.

  Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 20 20 30 30
Mean (pg/mL) 184 1890 191 1,867
Standard deviation 4.96 67 19.8 110
CV (%) 69.5 54.4 10.3 91.3
 

Recovery

Samples containing and/or spiked with human IL-17E were evaluated for recovery.

Sample Type Average % Recovery Range (%)
Cell culture supernate 120 109-138
Serum 55.2 25.8-81.0
EDTA plasma 58.8 41.7-79.0
Heparin plasma 59.5 47.9-73.8
Plaelet-poor EDTA 65.7 40.4-80.7
Platelet-poor Heparin 65.2 30.7-83.3
 

Linearity

Samples containing and/or spiked with human IL-17E were serially diluted to evaluate assay linearity.

  Cell culture supernate Serum EDTA plasma Heparin plasma Platelet-poor EDTA Platelet-poor Heparin
1:2 Average % of Expected 92.0 129 134 134 126 123
Range (%) 85.0-99.4 112-142 120-154 117-157 116-139 117-135
1:4 Average % of Expected 88.6 146 155 152 142 135  
Range (%) 79.9-98.0 117-178 128-194 128-199 134-163 126-149
1:8 Average % of Expected 86.5 161 174 167 154 141
Range (%) 79.2-94.3 122-205 133-242 134-236 139-185 128-170
1:16 Average % of Expected 84.4 161 178 167 156 141
Range (%) 77.0-92.6 122-216 129-264 130-245 140-196 124-183
Which Luminex Assay is right for me?

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.

More Information About Magnetic Bead-based Multiplex Kits For The Luminex® Platform

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

IL-17E/IL-25 XL Cytokine Luminex Panel Standard Curves

Background: IL-17E/IL-25

The IL-17 family is comprised of at least six proinflammatory cytokines that share a conserved cysteine-knot structure but diverge at the N-terminus. IL-17 family members are glycoproteins secreted as dimers that induce local cytokine production and recruit granulocytes to sites of inflammation. IL-17 is induced by IL-15 and IL-23, mainly in activated CD4+ T cells distinct from Th1 or Th2 cells. IL-17F is the most homologous to IL-17, but is induced only by IL-23 in activated monocytes. IL-17B binds the IL-17B receptor, but not the IL-17 receptor; it is most homologous with IL-17D, which is expressed by resting CD4+ T cells and CD19+ B cells. IL-17E is mainly produced by Th2 cells and recruits eosinophils to lung tissue. IL-17C has a very restricted expression pattern but has been detected in adult prostate and fetal kidney libraries.

Long Name
Interleukin 17E
Entrez Gene IDs
64806 (Human); 140806 (Mouse); 501996 (Rat)
Alternate Names
IL17E; IL-17E; IL25; IL-25; interleukin 25; Interleukin-17E; interleukin-25

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