HTRF HUMAN IL8 KIT-Revvity
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Cat.NO Name Size
62HIL08PEH HTRF HUMAN IL8 KIT 10K PTS

 

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

Also called CXCL8, IL8 is a chemokine mainly produced by macrophages, T cells, and neutrophils. IL8 acts as a chemoattractant neutrophils, and promotes infiltration and activation at inflammation sites. IL8 is involved in several cancer types due to 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.

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.

 

cytokines-how-it-works-assay-principle-human
 
 
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.

 

Revvity also worked with Myassays.com to help you in your data analysis.

 

 

Assay details

Technical specifications of human IL8 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

The same amount of recombinant cytokine was added to 2 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery found validates the sample matrix used for this assay.

 

 

Assay validation

Inhibition of IL8 secretion in THP1 cells with JTE607

THP1 cells plated at 100 k cells/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 IL8 Assay.

 

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IL8 secretion in PBMCs stimulated with LPS

PBMC plated at 50, 100, 200, and 400 k cells/well were stimulated for 3 h 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 IL8 Assay.

 

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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