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| Feature | Specification |
|---|---|
| Application | 細胞シグナル伝達 |
| Sample Volume | 10 µL |
DAP12, also known as TYRO protein (tyrosine kinase-binding protein), serves as a signaling adaptor for several cell surface receptors of the innate immune system. Crosslinking receptors related to DAP12 activate SYK or Zap-70 and other downstream targets, phosphorylate tyrosine residues in their cytoplasmic ITAM by SRC family kinases, and start downstream signaling. The TREM2-DAP12 receptor complex appears to be involved in maintaining a healthy brain homeostasis in microglia cells as well as monitoring the surroundings of neurons.
The AlphaLISA SureFire Ultra Mouse Total DAP12 Detection Kit is a sandwich immunoassay for the quantitative detection of total DAP12 in cellular lysates, using Alpha Technology.
The Total-AlphaLISA SureFire Ultra assay measures the expression level of a protein target in a cell lysate.
The Total-AlphaLISA SureFire Ultra assay uses two antibodies which recognize two different distal epitopes on the targeted protein. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the assay specific antibody, labeled with a CaptSure tag. Donor beads are coated with streptavidin to capture one of the detection antibodies, which is biotinylated. In the presence of targeted protein, the two antibodies bring the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of protein present in the sample.
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well OptiPlate™ plate before the addition of Total-AlphaLISA SureFire Ultra detection reagents. This protocol permits the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
Detection of Total target protein with AlphaLISA SureFire Ultra reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining AlphaLISA SureFire Ultra quality.
| Application |
Cell Signaling
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
AlphaLISA SureFire Ultra
|
| Detection Modality |
Alpha
|
| Lysis Buffer Compatibility |
Lysis Buffer
|
| Molecular Modification |
Total
|
| Product Group |
Kit
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
DAP12
|
| Target Class |
Phosphoproteins
|
| Target Species |
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Neuroscience
|
| Unit Size |
500 assay points
|
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