HTRF HUMAN TNF ALPHA KIT-Revvity
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Cat.NO Name Size
62HTNFAPEG HTRF HUMAN TNF ALPHA KIT 500 PTS
62HTNFAPEH HTRF HUMAN TNF ALPHA KIT 10K PTS

 

  • Product overview
  • Brand introduction
  • Q&A
  • Data download

TNF alpha is a cytokine mainly produced by macrophages, T cells, and neutrophils. TNF alpha acts as a chemoattractant neutrophils, and promotes infiltration and activation at inflammation sites. TNF alpha is involved in several cancer types 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 detection of low levels of analytes in serum or plasma. When assaying, always follow recommended protocol and avoid highly haemolyzed samples.

Specifications

 

 

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.

 

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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,  Revvity also worked with Myassays.com to help you in your data analysis. you’ll be able to access a free online software to run your TNF alpha analysis.

 

 

Assay details

Technical specifications of human TNF alpha kit

 

 

Analytical performance

Intra and inter assay

 

 

Dilutional linearity

​The excellent % of recovery obtained from these experiments show the good linearity of the assay.​

 

 

Spike and recovery

 

 

Assay validation

Inhibition of TNF alpha secretion in THP1 cells with JTE607

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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TNF alpha secretion in PBMCs stimulated with LPS

PBMC plated at 50, 100, 200 and 400 kcells/well were stimulated for 3h with increasing concentrations of LPS (0, 0.02, 0.2, 2 µg/mL). 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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At Revvity, we regard "impossibility" as inspiration and "inability" as the driving force. Revvity offers health science solutions, cutting - edge technologies and professional services, with its business covering the entire end - to - end process of scientific research exploration, development, diagnosis, and treatment. Relying on years of in - depth cultivation in fields such as translational multi - omics technology, biomarker identification, imaging, disease prediction, screening, detection and diagnosis, and informatics, Revvity is using the power of technology to break through the boundaries of human potential.

 

In 2023, Revvity's turnover exceeded $2.7 billion. It has more than 11,000 employees worldwide and provides diverse services for customers such as pharmaceutical and biotechnology enterprises, diagnostic laboratories, scientific research institutions and government agencies. The company is a member of the S&P 500 Index, and its customers are spread across more than 190 countries and regions around the world.

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