Human IL22 kit HTRF®

Name: Human IL22 kit
Brand: Cisbio
SKU: 44267
Availability: InStock

Associated products

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

No-wash
Highly specific
Under 2h bench time

Download the protocols

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

Size

Cat. number selected :

Quantity

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.

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/rat IL-22 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
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 size	16 µL
Final assay volume	20 µL
Kit components	Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD & LOQ (in Diluent)	9 pg/mL & 26 pg/mL
Range	26 – 5,000 pg/mL
Time to result	Overnight at RT (reading possible at 3h)
Calibration	No international standard available
Species	Human only

Intra and inter assay

Intra-assay (n=24)

Sample	Mean [IL22] (pg/mL)	CV
1	78	8%
2	411	5%
3	5000	6%
	Mean CV	6.3%

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
1	78	471	9.4%
2	411	2104	3.1%
3	5000	15181	1.7%
		Mean CV	4.7

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
1	4,000	2,094	2,159	103%
2	1,538	863	816	95%

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.

HTRF Product Catalog

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 Life Sciences 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

Protocol for Th17 differentiation from CD4 mouse T-lymphocyte

A step by step protocol for Th17 cells differentiation from naive CD4 mouse cells - Technical Notes

Safety Data Sheet (DEU) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (ELL) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (FRA-FR) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (ITA) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (SPA) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (ENG-GB) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (ENG-US) IL22 (h) Kit / 62HIL22PEG

62HIL22PEG - Safety Data Sheet

Safety Data Sheet (DEU) IL22 (h) Kit / 62HIL22PEH