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HTRF Human IL-4 Detection Kit, 500 Assay Points

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

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Feature	Specification
Application	Protein Quantification
Sample Volume	16 µL

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

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Product variants

Unit Size: 500 assay points

Part #:

62HIL04PEG

Unit Size: 10,000 assay points

Part #:

62HIL04PEH

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

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Product information

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 ir differentiation into plasma cells and ir class switch to immunoglobulin E, as well as promoting class II MHC expression. In immuno-oncology, IL4 induces 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 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 detection of low levels of analytes in serum or plasma.

How it works

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 Revvity 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.

Revvity also worked with Myassays.com to help you in your data analysis.

Assay details

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

Analytical performance

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

Specifications

Other specifications

Application	Protein Quantification
Brand	HTRF
Detection Modality	HTRF
Product Group	Kit
Sample Volume	16 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Cytokines
Target Species	Human
Technology	TR-FRET
Therapeutic Area	Oncology & Inflammation
Unit Size	500 assay points