Cynomolgus TNF alpha Assay Kit

TNFα, mainly produced by activated macrophages, is one of the main cytokines in acute phase reaction. Depending on the cellular target and the biological context, TNFα can activate pro-apoptosis or pro-survival pathways, thus regulating tissue development through apoptosis control.
Outside of Oncology and Inflammation, TNFα contributes to the development of Type II Diabetes by increasing insulin resistance and modulating fatty acid metabolism.
Cisbio's Cynomolgus TNFα kit is a homogeneous, add and read assay to measure TNFα secretion in supernatant. It offers a fast, efficient alternative to ELISA.

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
Download Insider tips for PBMC isolation Technical Note
Download cytokine release from fresh blood samples application notes

Assay principle

The assay protocol, using a Cisbio 96-well low volume or a 384-well small volume white plate (20 µL final), is described below. Cell supernatant, sample or standard are dispensed directly into the assay plate for the detection by HTRF® reagents. 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 in 96 to 1536-well plates by simply resizing each addition volume proportionally.

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.

Add

Cytokines assay principle add

Read

Cytokines assay principle read

(1) Assay data analysis:

  • "Guidelines for data reduction" button will show you how dealing with HTRF data processing and 4PL 1/y2 fitting.
  • "MyAssays" button will direct you to a free online software able to run TNFα analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

Intra-assay (n=24)

Sample Mean [TNFα] (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.

Inter-assay (n=4)

Sample [TNFα] (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.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Cynomolgus TNF alpha kit - 500 tests62CTNFAPEG
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Cynomolgus TNF alpha kit - 10,000 tests62CTNFAPEH
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Companion products

DescriptionCat. noProduct insertMSDS
Cynomolgus TNF alpha kit - 3 Standards62CTNFACD3
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Cynomolgus TNF alpha kit - Standard62CTNFACDA
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