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| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Dynamic Range | 5.1 - 300,000 pg/mL |
| Limit of Detection | 5.1 pg/mL |
| Limit of Quantification | 16.7 pg/mL |
| Sample Volume | 5 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.
Tumor necrosis factor alpha (TNFα) is a multifunctional proinflammatory cytokine produced primarily by activated macrophages and T cells. TNFα is primarily produced as 233 Amino-Acid long TNF type II transmembrane proteins. The soluble form of TNFα can be generated from its transmembrane form via proteolytic cleavage by ADAM17. Released TNFα self-associates to form a soluble complex that exerts its function by binding the TNFR1 and TNFR2 receptors. TNFα has been observed to play a role in inflammation, apoptotic cell death, and as an important regulator of tumorigenesis and infection. TNFα dysregulation has been observed in several major diseases such as Inflammatory Bowel Disease, Depression, Cancer, and Neurological Diseases. This kit has been designed for the detection of TNFα in cynomolgus and rhesus monkey serum and cell culture supernatants.
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 |
5.1 - 300,000 pg/mL
|
| Limit of Detection |
5.1 pg/mL
|
| Limit of Quantification |
16.7 pg/mL
|
| Product Group |
Kit
|
| Sample Volume |
5 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
TNFα
|
| Target Class |
Cytokines
|
| Target Species |
Cynomolgus
|
| Technology |
Alpha
|
| Therapeutic Area |
Inflammation
|
| Unit Size |
5,000 assay points
|
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