| Feature | Specification |
|---|---|
| Application | 細胞シグナル伝達 |
| Protocol Time | 2h at RT |
| Sample Volume | 30 µL |
Tau aggregates are pathological assemblies of hyperphosphorylated tau protein, commonly observed in tauopathies such as Alzheimer's disease (AD), frontotemporal dementia (FTD), and progressive supranuclear palsy (PSP). Tau is a microtubule-associated protein that stabilizes axonal microtubules under normal conditions. In disease states, abnormal post-translational modifications such as hyperphosphorylation cause tau to detach from microtubules and misfold into toxic oligomers and fibrils, forming neurofibrillary tangles. These aggregates disrupt neuronal transport, synaptic integrity, and contribute to neuroinflammation and cell death. Therapeutic efforts aim to inhibit Tau aggregation, enhance its clearance, or block its spread between neurons.
The AlphaLISA SureFire Ultra Human Tau Aggregate Detection Kit is a sandwich immunoassay for the quantitative detection of Tau Aggregate in cellular lysates, using Alpha Technology.
Formats:
AlphaLISA SureFire Ultra kits are compatible with:
AlphaLISA SureFire Ultra kits can be used for:
The Aggregation-AlphaLISA SureFire Ultra assay measures the aggregates level of a protein target in a cell lysate.
The Aggregation-AlphaLISA Surefire Ultra assay uses one antibody which recognizes an exposed epitope on the aggregated target 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 the detection antibody, which is biotinylated. In the presence of targeted protein, the antibody brings the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, enabling the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of aggregated 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 Aggregation-AlphaLISA SureFire Ultra detection reagents. This protocol enables the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
Detection of aggregated 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 enables miniaturization while maintaining robust AlphaLISA SureFire Ultra quality.
Tau Aggregate assay was assessed by assaying recombinant aggregated protein. Serial dilutions of recombinant Human Tau aggregate protein (Abcam ab246003) was prepared beginning at 100 ng/mL in Lysis Buffer and evaluated using the Tau Aggregate AlphaLISA SureFire Ultra assay.
For the detection step, 10 µL of protein was transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor mix and incubated for 1 hour at room temperature. Finally, 5 µL of Donor mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision using standard AlphaLISA settings.
| Application |
Cell Signaling
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
AlphaLISA SureFire Ultra
|
| Detection Modality |
Alpha
|
| Product Group |
Kit
|
| Protocol Time |
2h at RT
|
| Sample Volume |
30 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
Tau
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Neuroscience
|
| Unit Size |
100 assay points
|
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