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| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Dynamic Range | 34 - 30,000 pg/mL |
| Limit of Detection | 15 pg/mL |
| Limit of Quantification | 34 pg/mL |
| Sample Volume | 5 µL |
Insulin is produced in the pancreas by Beta-cells of the Langherans islets as a 110 amino acid proinsulin hormone. Proinsulin is post-translationally cleaved into two chains (peptide A of 21 amino acids and peptide B of 30 amino acids) that are covalently bonded through two disulfide bonds and released when blood glucose levels rise. After 30 minutes, the blood concentration rises from roughly 50 pmol/L to 300-400 pmol/L. Insulin regulates glucose and lipid metabolism and has been linked to several disorders, including diabetes, heart disease, and obesity.
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 |
34 - 30,000 pg/mL
|
| Limit of Detection |
15 pg/mL
|
| Limit of Quantification |
34 pg/mL
|
| Product Group |
Kit
|
| Sample Volume |
5 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
Insulin
|
| Target Class |
Biomarkers
|
| Target Species |
Human
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Metabolic
|
| Unit Size |
5,000 assay points
|
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