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