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| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Dynamic Range | 4.5 - 100,000 pg/mL |
| Limit of Detection | 4.5 pg/mL |
| Sample Volume | 5 µL |
Formats:
Features:
Human Interleukin 5 (IL5) is produced as a precursor protein maturing into a 115 amino acid protein. The mature active protein is heavily glycosylated and is covalently linked in a homodimer by a disulfide bond, with a native apparent molecular weight of 45-60 kDa. It is produced by Th2 cells, mast cells, and eosinophils. IL5 stimulates B cell growth and promotes the production of cytotoxic T-cells from thymocytes, but its key function is to mediate activation, maturation, and survival of eosinophils. IL5 activated eosinophils eliminate antibody bound parasites through the release of cytotoxic granule proteins. IL5 seems to play a major role in the development of allergic disease/asthma response, which makes IL5 an interesting target for the development of anti-allergy drugs. IL5 is also a potential marker of acute graft-versus-host disease.
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 |
4.5 - 100,000 pg/mL
|
| Limit of Detection |
4.5 pg/mL
|
| Product Group |
Kit
|
| Sample Volume |
5 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
IL5
|
| Target Class |
Cytokines
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Inflammation
|
| Unit Size |
500 assay points
|
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