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| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Dynamic Range | 42 - 100,000 pg/mL |
| Limit of Detection | 42 pg/mL |
| Limit of Quantification | 104 pg/mL |
| Sample Volume | 10 µL |
Formats:
Features:
AlphaLISA technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In an AlphaLISA assay, a Biotinylated Anti-Analyte Antibody binds to the Streptavidin-coated Alpha 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 provokes 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.
CD276 (Cluster of Differentiation 276, or B7-H3) is a type I transmembrane protein belonging to the B7 family of co-stimulatory proteins (e.g., B7-1, or CD80). It signals through CD28 family molecules such as CD28 and CTLA4. It is expressed ubiquitously, such as dendritic cells derived from monocytes, epithelial cells of sinonasal tissue, extravillous trophoblast cells and Hofbauer cells of the placenta. It is an immune checkpoint molecule, involved in the regulation of T-cell-mediated immune response. It is expressed by some solid tumours, as a biomarker for detection of neuroblastoma cells and is the target of anticancer therapy.
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 |
42 - 100,000 pg/mL
|
| Limit of Detection |
42 pg/mL
|
| Limit of Quantification |
104 pg/mL
|
| Product Group |
Kit
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
CD276
|
| Target Class |
Biomarkers
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Oncology
|
| Unit Size |
100 assay points
|
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