Cynomolgus TNF alpha kit HTRF®

The HTRF cynomolgus TNF alpha kit is intended for the quantification of cynomolgus TNF alpha release in cell supernatant.

No-wash
Low sample consumption
Under 2h bench time

The HTRF cynomolgus TNF alpha kit is intended for the quantification of cynomolgus TNF alpha release in cell supernatant.

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Overview

TNF alpha, mainly produced by activated macrophages, is one of the main cytokines in acute phase reaction. Depending on the cellular target and the biological context, TNF alpha can activate pro-apoptosis or pro-survival pathways, thus regulating tissue development through apoptosis control. Outside of oncology and inflammation, TNF alpha contributes to the development of type II diabetes by increasing insulin resistance and modulating fatty acid metabolism.

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 Cynomolgus TNFα 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
PANEL QUANTIFICATION

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 TNF alpha analysis.

Technical specifications of Cynomolgus TNF alpha

Sample size	16 µL
Final assay volume	20 µL
Kit components	Lyophilized standard, frozen detection antibodies, buffers & protocol.
LOD & LOQ (in Diluent)	12.4 pg/mL & 27 pg/mL
Range	27 – 3,000 pg/mL
Time to result	ON at RT
Species	Cynomolgus only

Inter and Intra assay

Sample	Mean [TNFa] (pg/mL)	CV
1	100	14%
2	1500	2%
3	2750	2%
	Mean CV	6%

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

Sample	[TNFa] (pg/mL)	Mean (delta R)	CV
1	159	665	3%
2	514	2150	2%
3	1667	6467	2%
		Mean CV	2%

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

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