Human CXCL5/ENA-78 Magnetic Luminex® Performance Assay
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Human CXCL5/ENA-78 Magnetic Luminex® Performance Assay Summary
| Panel | Luminex Performance Human Cytokine Panel A |
| 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. |
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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 CXCL5/ENA-78 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
- 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 Base Kit.
- User-mix antibody-coated Magnetic beads in individual vials
- User-mixed cocktail of biotinylated detection antibody
Precision
|
Intra-assay Precision (precision within an assay) - Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. |
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| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 296 | 682 | 1446 | 310 | 682 | 1390 |
| Standard deviation | 14 | 28 | 44.2 | 33 | 69 | 127 |
| CV (%) | 4.7 | 4.1 | 3.1 | 10.6 | 10.1 | 9.1 |
Recovery
|
Samples containing and/or spiked with human ENA-78 were evaluated for recovery. |
||||||
| Sample Type | Average % Recovery | Range (%) | ||||
|---|---|---|---|---|---|---|
| Cell culture supernate | 105 | 92-118 | ||||
| Serum | 97 | 79-122 | ||||
| EDTA plasma | 102 | 86-121 | ||||
| Heparin plasma | 108 | 96-126 | ||||
Linearity
|
Samples containing and/or spiked with human ENA-78 were serially diluted to evaluate assay linearity. |
|||||||||
| Cell culture supernate | Serum | EDTA plasma | Heparin plasma | ||||||
|---|---|---|---|---|---|---|---|---|---|
| 1:2 | Average % of Expected | 95 | 100 | 100 | 99 | ||||
| Range (%) | 86-119 | 87-106 | 87-139 | 85-107 | |||||
| 1:4 | Average % of Expected | 89 | 100 | 95 | 97 | ||||
| Range (%) | 85-95 | 90-107 | 49-170 | 79-110 | |||||
| 1:8 | Average % of Expected | 88 | 99 | 97 | 91 | ||||
| Range (%) | 83-96 | 86-115 | 67-188 | 71-104 | |||||
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
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Quantist software graph of signal intensity versus concentration for CXCL5/ENA-78 in the Human Cytokine A Luminex Panel
Background: CXCL5/ENA-78
CXCL5/Epithelial Cell-derived Neutrophil-activating Peptide (ENA-78), is a member of the CXC subfamily of chemokines. Full-length CXCL5/ENA-78 is 114 amino acids (aa) in length with a predicted molecular weight of 12 kDa. Following the removal of the signal peptide, bioactive CXCL5/ENA-78 is 78 aa in length. CXCL5/ENA-78 can be N-terminally cleaved by Cathepsin G and Chymotrypsin to CXCL5/ENA-74 (74 aa) and CXCL5/ENA-70 (70 aa), which show increased potency relative to CXCL5/ENA-78. While murine LIX was thought to be an ortholog to human CXCL5/ENA-78, genome-wide analysis and a consensus in the field suggests that human CXCL5/ENA-78 does not have a true murine ortholog. CXCL5/ENA-78 is upregulated at sites of inflammation and is expressed by multiple hematopoietic cell types, fibroblasts, endothelial cells, vascular smooth muscle cells, and adipocytes. It acts as a chemoattractant for neutrophils, has angiogenic properties, and contributes to cancer progression.
Citation for Human CXCL5/ENA-78 Magnetic Luminex® Performance Assay
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
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Patients with idiopathic recurrent miscarriage have abnormally high TGF�+ blood NK, NKT and T cells in the presence of abnormally low TGF� plasma levels
Authors: L Zhu, M Aly, RJ Kuon, B Toth, H Wang, H Karakizlis, R Weimer, C Morath, E Ibrahim, N Ekpoom, G Opelz, V Daniel
BMC Immunol., 2019-03-04;20(1):10.
Species: Human
Sample Types: Plasma
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