
Feature | Specification |
---|---|
Application | Cell Signaling |
Sample Volume | 30 µL |
Receptor-interacting serine/threonine-protein kinase 3 (RIPK3) is a critical regulator of programmed necroptosis, an inflammatory form of cell death with important functions in pathogen-induced and sterile inflammation. RIP3 mediates necroptosis in conjunction with RIPK1 and MLKL. Due to the inflammation caused by necroptosis, RIPK3 is associated with a variety of inflammatory diseases.
The AlphaLISA SureFire Ultra Human Phospho-RIP3 (Ser227) assay is a sandwich immunoassay for the quantitative detection of phospho-RIP3 (Ser227) in cellular lysates, using Alpha Technology.
The Phospho-AlphaLISA SureFire Ultra assay measures a protein target when phosphorylated at a specific residue.
The assay uses two antibodies which recognize the phospho epitope and a distal epitope 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 phosphorylated 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 phosphoprotein 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 Phospho-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 Phosphorylated 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 |
Phosphorylation
|
Product Group |
Kit
|
Sample Volume |
30 µL
|
Shipping Conditions |
Shipped in Blue Ice
|
Target |
RIP3
|
Target Class |
Phosphoproteins
|
Target Species |
Human
|
Technology |
Alpha
|
Therapeutic Area |
Autoimmunity
Central Nervous System
Neuroinflammation
|
Unit Size |
100 assay points
|
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