Human IL4 kit HTRF®

Name: Human IL4 kit
Brand: Cisbio
SKU: 40415
Availability: InStock

Associated products

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

No-wash
Low sample consumption
Under 2h bench time

Download the protocols

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.

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

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