The Human IL4 kit is designed for the quantification of human IL4 release in cell supernatant.
See more
  • No-wash
  • Low sample consumption
  • Under 2h bench time
The Human IL4 kit is designed for the quantification of human IL4 release in cell supernatant.
-
In stock

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.

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 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

Normalize HTRF cytokine assays with viability assay on Spectramax I3X-Minimax

In collaboration with Molecular Devices - Application Notes

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

Validated protocol - 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

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

tab6