No-wash kit to quantify released Human IFN gamma
Human TNF alpha kit
This HTRF kit is designed for the quantification of human TNF alpha release in cell supernatant.
- Validated on PBMC
- Under 2h bench time
TNF alpha is a cytokine mainly produced by macrophages, T cells, and neutrophils. TNF alpha acts as a chemoattractant for neutrophils, and promotes infiltration and activation at inflammation sites. TNF alpha is involved in several cancer types for its ability to promote angiogenesis and cell proliferation, as well as to inhibit apoptosis.
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.
|Sample size||16 µL|
|Final assay volume||20 µL|
|Kit components||Lyophilized standard, frozen detection antibodies, buffers &protocol.|
|LOD &LOQ (in Diluent)||5 pg/mL &29 pg/mL|
|Range||29 2,500 pg/mL|
|Time to result||2h at RT|
|Calibration||NIBSC (12/154) value (IU/mL) = 0,1 x HTRF hTNFa value (pg/mL)|
|Sample||Mean [TNFα] (pg/mL)||CV|
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
|Sample||[TNFa] (pg/mL)||Mean (delta R)||CV|
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
The excellent % of recovery obtained from these experiments show the good linearity of the assay.
|Dilution factor||[TNFα] expected (pg/mL)||[TNFα] detected (pg/mL)||Recovery|
|[TNFα] added (pg/mL)||[TNFα] expected (pg/mL)||[TNFα] detected (pg/mL)||Recovery|
THP1 cells plated at 100 kcells/well were incubated with increasing concentrations of JTE 607 for 18h, then stimulated for 3 h with 2 µg/mL LPS. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human TNFa Assay.
Can HTRF technology answer to different scientific questions? An academic with industrial standards point of view
In collaboration with CNRS - Scientific Presentations
In collaboration with GSK - Scientific Presentations
In collaboration with Molecular Devices - Application Notes
In collaboration with Blood assay solution - Application Notes
Perform and optimize cytokine assays on PBMC - Technical Notes
Molecular Devices - Application Notes
A fun video introducing you to cytokines assays with HTRF - Videos
cGAS-STING signaling pathway from A to Z - Brochures
For researcher working on checkpoint inhibitors - Application Notes
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