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| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Dynamic Range | 73 - 300,000 pg/mL |
| Limit of Detection | 73 pg/mL |
| Limit of Quantification | 234 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.
Most mammal IgG class antibodies, including those from human are composed of two heavy and two light chains. Such light chains can be of kappa or lambda types, with an expected ratio of 3 kappas to one lambda. Both appear to play the same function in antibodies. Free light chains are usually rare in blood. However, several types of multiple myelomas will secrete free light chains, which can then accumulate in organs such as the kidney and cause severe damage, with lambda chains being the poorest diagnostic.
Knowledge of the type of light chain generated is also important for the creation and production of therapeutic antibodies. Techniques used for purification, such as protein L or anti species antibodies, can have different affinities for the two chains. The kappa light chain AlphaLISA detection kit allows for the detection of kappa light chain in human sera, plasma, 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 |
73 - 300,000 pg/mL
|
| Limit of Detection |
73 pg/mL
|
| Limit of Quantification |
234 pg/mL
|
| Product Group |
Kit
|
| Sample Volume |
5 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
Kappa light chain
|
| Target Class |
Biologics
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Unit Size |
5,000 assay points
|
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