Description
Formats:
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
Intercellular Adhesion Molecule-1 (ICAM-1) is a dimeric transmembrane glycoprotein with a molecular weight of 80 - 114 kDa. ICAM-1 is a member of the immunoglobulin family and can be expressed on non-hematopoietic cells, such as vascular endothelial cells. It is involved in the transmigration of leucocytes to sites of inflammation. A soluble form of ICAM-1 (sICAM-1) has been identified in serum and cerebrospinal fluid and it contains the five extracellular Ig-domains of ICAM-1. sICAM-1 is an inflammatory marker that has been associated with several common diseases such as diabetes, heart disease, stroke, and malaria. High levels of sICAM-1 have been found in serum from patients with cardiovascular disease, cancer, HIV-1, and autoimmune diseases. sICAM-1 is able to competitively bind the ligands of membrane-bound ICAM-1, such as LFA-1, Mac-1, and human rhinovirus.
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 assay
The AlphaLISA assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-analyte antibody bound to Streptavidin-coated 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 assay
The AlphaLISA 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.
