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HTRF Human IL-13 Detection Kit, 10,000 Assay Points
The HTRF human IL13 kit is designed for the quantification of human IL13 release in cell supernatant.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Product Variants
Product information
Overview
Interleukin 13 plays an essential role in allergic inflammation, airway disease, and asthma by regulating IgE synsis and inducing B-cell antibody class switching. IL13 can also induce differentiation of naive T-cells into Th2. Its effect on immune cells is similar to IL4 (close secondary structures). Outside immune cells, IL13 can induce matrix metalloproesases (MMPs) in fibroblasts as a mechanism to avoid asphixation during excessive allergic inflammation. Human IL13 is produced by Th2, CD4+, Tregs, NK, mast cells, basophils, and eosinophils, as well as epilial cells. After being secreted, it binds to IL13Ra1, which complexes with IL4Ra to form type II IL4R heterocomplex. This leads to activation of Jak1, Tyk2, and downstream signaling through STAT3/6.
HTRF human IL13 kit is a homogeneous, add and read assay to measure IL-13 secretion in cell supernatants. It offers a fast, efficient alternative to ELISA.
Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable detection of low levels of analytes in serum or plasma.
Specifications
| Assay Points |
10000
|
|---|---|
| Assay Target Type |
Kit
|
| Assay Technology |
HTRF
|
| Brand |
HTRF
|
| Quantity |
1
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Therapeutic Area |
Inflammation
|
| Unit Size |
10,000 Assay Points
|
Video gallery
How it works
Assay Principle
Cell supernatant, sample, or standard is dispensed directly into the assay plate for detection by HTRF® reagents (384-well low-volume white plate or Revvity low-volume 96-well plate in 20 µL). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.
Assay data analysis
The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.
To fully understand how to deal with HTRF data processing, and also 4PL 1/y² fitting,
Revvity also works to help you in your data analysis.
Assay details
Technical specifications of human IL13 kit
| Sample size | 16 µL |
|---|---|
| Final assay volume | 20 µL |
| Kit components | Lyophilized standard, frozen detection antibodies, buffers, & protocol. |
| LOD & LOQ (in Diluent) | 6 pg/mL & 17 pg/mL |
| Range | 17 – 5,000 pg/mL |
| Time to result | Overnight at RT |
| Calibration | NIBSC (94/622) value (IU/mL) = 0.0012 x HTRF hIL13 value (pg/mL) |
| Species | Human only |
Analytical performance
Intra and inter assay
Intra-assay (n=24)
| Sample | Mean [IL13] (pg/mL) | CV |
|---|---|---|
| 1 | 50 | 8% |
| 2 | 2500 | 10% |
| 3 | 4000 | 9% |
| Mean CV | 9% |
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
Inter-assay (n=4)
| Sample | [IL13] (pg/mL) | Mean (delta R) | CV |
|---|---|---|---|
| 1 | 78 | 685 | 9% |
| 2 | 411 |
2780 | 5% |
| 3 | 5000 | 13729 | 3% |
| Mean CV | 6% |
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
Spike and recovery
| Sample | [IL13] added (pg/mL) | [IL13] expected (pg/mL) | [IL13] detected (pg/mL) | Recovery |
|---|---|---|---|---|
| 1 | 945 | 1,307 | 1,181 | 90% |
| 2 | 2,174 | 1,922 | 1,728 | 90% |
Two different amounts of recombinant IL13 cytokine were added to RPMI + 10% FCS cell supernatant samples containing a known value of IL13 (1669 pg/mL). Samples were mixed at a 1:1 volume ratio. The set of responses obtained from a standard curve was compared to the calculated expected values. The recovery value observed validates the sample matrix used for this assay.
Assay validation
IL13 secretion in PBMCs stimulated with PHA
Human PBMCs were plated at 100 kcells/well in a 96-well plate under 100 µL of complete RPMI cell culture medium, then stimulated with increasing concentrations of PHA (0,4 to 50 µg/mL). After a 3-day incubation, 16 µL of supernatants were transferred into a white detection plate to be analyzed with the Human IL13 Assay.
Resources
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SDS, COAs, Manuals and more
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