Description
Formats:
Our HV (high volume) 100 assay point kit allows you to run 100 wells in 96-well format, using a 100 μL reaction volume (10 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
Half the time of an ELISA assay
Interleukin-17A (IL-17A) is the founding member of a group of cytokines called the IL-17 family. The other members are IL-17B, C, D, E, and F. Mature mouse or rat IL-17A is a glycosylated homodimer formed of two 133 amino acid subunits. IL-17A is produced by a subset of T helper cells. It is a proinflammatory cytokine that enhances T cell priming and stimulates macrophages, fibroblasts, endothelial and epithelial cells to produce mediators of inflammation like IL-1, IL-6, and TNFα. It is also critical for neutrophil activation and migration, and induces IL-8, a key chemokine for neutrophils. IL-17 has been involved in the proinflammatory patterns associated with joint inflammation and rheumatoid arthritis in mouse and human models and may be a novel therapeutic target for the treatment of these diseases.
AlphaLISA™ technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA biotin-free assay, a DIG-labeled anti-analyte antibody binds to the anti-DIG-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 biotin-free assay
The AlphaLISA Biotin-Free assay is based on an AlphaLISA sandwich immunoassay involving a digoxigenin anti-analyte antibody bound to anti-DIG AlphaLISA Donor beads and an anti-analyte antibody conjugated to AlphaLISA Acceptor beads. Both antibodies are directed against the analyte of interest. In the presence of the target, both antibodies bind to analyte 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 analyte present in the sample.
Protocol of the AlphaLISA biotin-free assay
The AlphaLISA Biotin-Free assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples or standards are dispensed directly into the assay plate for the detection of the analyte of interest by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
