Human IFN gamma kit
No-wash kit to quantify released Human IFN gamma
The HTRF human TNFα kit is designed for the quantification of human TNF alpha release in cell supernatant.
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.
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 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.
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.
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Molecular Devices - Application Notes
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