| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Dynamic Range | 7 - 100,000 pg/mL |
| Limit of Detection | 7 pg/mL |
| Limit of Quantification | 24 pg/mL |
| Sample Volume | 10 µL |
Formats:
Features and Benefits:
Tumor Necrosis Factor Alpha is a multifunctional proinflammatory cytokine synthesized mainly by nucleated blood cells as a 233 aa type II transmembrane protein which is cleaved by ADAM17 between aa76-77 to form a soluble homotrimeric complex. TNFα plays a role in lipid metabolism, coagulation and endothelial function and has been associated with cancer, infection and inflammation (including inflammatory bowel disease), ischemia/reperfusion injury and heart failure, and insulin resistance. This kit is used when the analyte is in a matrix containing high levels of biotin, such as can be found in cell culture media such as RPMI, or milk.
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA biotin-free assay, a DIG-labeled anti-analyte antibody binds to the anti-DIG-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 Biotin-Free assay is based on an AlphaLISA sandwich immunoassay involving a digoxigenin anti-analyte antibody bound to anti-DIG 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 Biotin-Free 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 |
7 - 100,000 pg/mL
|
| Limit of Detection |
7 pg/mL
|
| Limit of Quantification |
24 pg/mL
|
| Product Group |
Kit
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
TNFα
|
| Target Class |
Cytokines
|
| Target Species |
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Inflammation
|
| Unit Size |
100 assay points
|
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AlphaLISA™ technology is a highly sensitive, easy-to-use, and reproducible method for detecting and quantifying molecules in...
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