
Feature | Specification |
---|---|
Application | Protein Quantification |
Dynamic Range | 7.3 - 1,000,000 pg/mL |
Limit of Detection | 7.3 pg/mL |
Sample Volume | 5 µL |
Formats:
Features:
C-reactive Protein (CRP) is an acute-phase serum protein (206 amino acids), present in homopentameric form, that plays a role in the first line of host defense. In response to infection, inflammation, or tissue damage, IL6, IL1, and glucocorticoids induce the production of CRP in the liver. CRP exhibits Ca2+-dependent binding to ligands such as phosphorylcholine, a constituent of many bacterial and fungal cell walls, or the membrane of injured cells. It also binds to nuclear components of necrotic and apoptotic cells. It has been shown that high levels of CRP in humans are associated with an increased risk of cardiovascular diseases. Its mechanism is not yet completely understood.
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.
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.
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.
Application |
Protein Quantification
|
---|---|
Automation Compatible |
Yes
|
Brand |
AlphaLISA
|
Detection Modality |
Alpha
|
Dynamic Range |
7.3 - 1,000,000 pg/mL
|
Limit of Detection |
7.3 pg/mL
|
Product Group |
Kit
|
Sample Volume |
5 µL
|
Shipping Conditions |
Shipped in Blue Ice
|
Target |
CRP
|
Target Class |
Biomarkers
|
Target Species |
Human
|
Technology |
Alpha
|
Therapeutic Area |
Inflammation
|
Unit Size |
5,000 assay points
|
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