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Human IL4 kit HTRF®

The HTRF human IL4 kit is designed for the quantification of human IL4 release in cell supernatant.

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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF human IL4 kit is designed for the quantification of human IL4 release in cell supernatant.

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Overview

IL4 is expressed by T cells, NK cells, and mast cells, and is an important regulator of humoral and adaptative immunity. It stimulates activated B cells, and induces their differentiation into plasma cells and their class switch to immunoglobulin E, as well as promoting class II MHC expression. In immuno-oncology, IL4 induces the differentiation of naïve T cells into TH2 cells, which in turn produce IL4 and thus create a positive feedback loop. IL4 is also involved in the development of airway inflammation and allergic asthma.


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-4 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

  • SCALABLE FROM 96 TO 1536WELLS
  • TH2 CELLS ACTIVATION READOUT

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.
IL4 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 worked 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 IL4 analysis.

Technical specifications of human IL4 kit

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent) 8 pg/mL &29 pg/mL
Range 29 – 4,000 pg/mL
Time to result 2h at RT
Calibration NIBSC (88/656) value (IU/mL) = 0,012 x HTRF hIL4 value (pg/mL)
Species Human only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [IL4] (pg/mL) CV
1 50 7%
2 1895 2%
3 3598 3%
Mean CV 4%

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

Inter-assay (n=4)

Sample [IL4] (pg/mL) Mean (delta R) CV
1 98 527 12%
2 432 2061 7%
3 1905 7265 6%
Mean CV 8%

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

HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

Inflammation cell by cell

HTRF solutions for each cell type - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the Immune System - Brochures

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Application Notes

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Technical Notes

How to freeze PBMC

Guideline for PBMC freezing - Technical Notes

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

Product Insert IL4 (h) Kit / 62HIL04PEG-62HIL04PEH

62HIL04PEG-62HIL04PEH - Product Insert

HTRF makes cytokine detection easy

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

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

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

Innovative solutions in Virology research

Kits and reagents for virology research - Flyers

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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