Description
IL6 is a pro-inflammatory cytokine involved in acute-phase reaction, inflammation, and cancer progression. Secreted by T cells, macrophages, and fibroblasts, it induces B and T cell proliferation. It is being studied in a wide variety of research areas including diabetes, Alzheimer's disease, depression, and several cancers. Along with TGF beta, IL6 is main promoter of T cell differentiation into TH17, a new component of immuno-oncology. AlphaLISA assays may have sufficient sensitivity to enable detection of low levels of analytes in serum or plasma.
Formats
Our 100 assay point kit allows you to run 100 wells in 96-well format, using a 100 µL reaction volume (5 µL of sample).
Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Features
No-wash steps, no separation steps
ELISA alternative technology
Sensitive detection
Broad sample compatibility
Small sample volume
Results in less than 3 hours
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Principle of the AlphaLISA HP human IL-6 assay
The AlphaLISA HP IL-6 assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-IL-6 antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-IL-6 antibody conjugated to AlphaLISA Acceptor beads. Both antibodies are directed against the IL-6 protein. In the presence of the target, both antibodies bind to IL-6 and the beads come into proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer within the Acceptor beads, resulting in emission with λmax at 615 nm. The intensity of the signal is directly proportional to the concentration of IL-6 present in the sample.
Protocol of the AlphaLISA HP human IL-6 assay
The AlphaLISA HP Human IL-6 assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples (cell supernatants or serum) or standards are dispensed directly into the assay plate for the detection of IL-6 by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
