Description
Formats:
Our 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
Chemokine ligand 4 (CXCL4), otherwise known as platelet factor 4 (PF4) is a small 8 kDa cytokine belonging to the CXC chemokine family. During platelet aggregation, CXCL4/PF4 is released from alpha-granules of activated platelets. The active protein is a tetramer ring of PF4 subunits which binds with high affinity to heparin. The function of CXCL4/PF4 is to promote blood clotting by neutralization of the anticoagulant effect of heparin molecules, by binding them on the endothelial surface of blood vessels. Mature human CXCL4/PF4 shares 96% amino acid sequence identity with the variant CXCL4L1/PF4V and 75% with mouse CXCL4/PF4. During heparin administration, some patients generate antibodies against the CXCL4/PF4/heparin complex, which activate platelets and can cause heparin-induced thrombocytopenia (HIT) and/or arterial thrombosis. The CXCL4/PF4 AlphaLISA detection kit allows for the detection of CXCL4/PF4 in human serum, plasma, and cell culture supernatants.
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.
