The HTRF human IL13 kit is designed for the quantification of human IL13 release in cell supernatant.

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  • No-wash No-wash
  • Highly specific Highly specific
  • Under 2h bench time Under 2h bench time

The HTRF human IL13 kit is designed for the quantification of human IL13 release in cell supernatant.

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Overview

Interleukin 13 plays an essential role in allergic inflammation, airway disease, and asthma by regulating IgE synthesis and inducing B-cell antibody class switching. IL13 can also induce the 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 epithelial cells. After being secreted, it binds to IL13Ra1, which complexes with IL4Ra to form the 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 the detection of low levels of analytes in serum or plasma. Check out the Human IL-13 alpha AlphaLISA kit’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples.

Benefits

  • VALIDATED ON PBMC
  • SHORT TIME-TO-RESULT

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 Cisbio 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.

IL13 kit assay principle

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, please visit this page.

Cisbio also works with Myassays.com to help you in your data analysis. By clicking on this link, you’ll be able to access a free online software to run your IL13 analysis.

Technical specifications of human IFN gamma kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers, & protocol.
LOD & LOQ (in Diluent)6 pg/mL & 17 pg/mL
Range 17 – 5,000 pg/mL
Time to resultOvernight at RT
CalibrationNIBSC (94/622) value (IU/mL) = 0.0012 x HTRF hIL13 value (pg/mL)
SpeciesHuman only

Intra and inter assay

Intra-assay (n=24)

SampleMean [IL13] (pg/mL)CV
1508%
2250010%
340009%

Mean CV9%

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
1786859%
2411
27805%
35000137293%


Mean CV6%

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
19451,3071,18190%
22,1741,9221,72890%

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.

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.

Illstration of the IL13 secretion in PBMCs stimulated with PHA
Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

HTRF makes cytokine detection easy

A fun video introducing you to cytokines assays with HTRF - Videos

Product Insert IL13 (h) Kit / 62HIL13PEG-62HIL13PEH

62HIL13PEG-62HIL13PEH - Product Insert

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

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