This kit is designed for the quantification of human IL22 release in cell supernatant.

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

This kit is designed for the quantification of human IL22 release in cell supernatant.

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In stock

Overview

Interleukin 22 is a type 2 cytokine (member of the IL10 family) and plays an essential role in innate protective immunity against extracellular pathogens. It induces pro-inflammatory responses, allows production of antimicrobial peptides. IL-22 is also involved in wound-healing and regeneration of epithelial cells. Human IL-22 is produced on inflammation site by Th17 CD4+ cells, activated T-cells, Th1 and NK cells. After being secreted, it targets non-hematopoietic cells and binds to IL22R that is restricted to tissue-resident cells (fibroblasts, epithelial cells). Downstream signaling involves phosphorylation of Jak1 and Tyk2 and subsequent activation of STAT3. Cisbio’s Human IL22 kit is a homogeneous, add and read assay to measure IL-22 secretion in cell supernatants. It offers a fast, efficient alternative to ELISA.

Benefits

  • VALIDATED ON PBMC
  • CORRELATED TO ELISA

Assay Principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the 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.

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 IL22 analysis.

Technical specifications of human IL22 kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers &protocol.
LOD & LOQ (in Diluent)9 pg/mL & 26 pg/mL
Range26 – 5,000 pg/mL
Time to resultOvernight at RT (reading possible at 3h)
CalibrationNo international standard available
SpeciesHuman only

Intra and inter assay

Intra-assay (n=24)

SampleMean [IL22] (pg/mL)CV
1788%
24115%
350006%

Mean CV6.3%

Each of the 3 samples was measured 24 times, and %CV for concentration was calculated for each sample.

Inter-assay (n=4)

Sample[IL22] (pg/mL)Mean (delta R)CV
178471
9.4%
24112104
3.1%
3500015181
1.7%


Mean CV4.7

Each of the samples was measured in 4 different experiments, and %CV for concentration was calculated for each sample.

Spike and recovery

Sample


[IL22] added (pg/mL)[IL22] expected (pg/mL)[IL22] detected (pg/mL)Recovery
14,0002,0942,159103%
21,53886381695%

Two different amounts of recombinant IL22 cytokine were added to RPMI + 10% FCS cell supernatant samples containing a known value of IL22 (188 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.

IL22 secretion in PBMCs stimulated with PHA

Human PBMCs were plated at 800 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 100 µg/mL). After a 3-day incubation, 16 µL of supernatants were transferred into a white detection plate to be analyzed with the Human IL22 Assay.

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 IL22 (h) Kit / 62HIL22PEG-62HIL22PEH

62HIL22PEG-62HIL22PEH - 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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