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The AlphaLISA™ SureFire® Ultra™ Human and Mouse Total Aggregate TDP-43 assay is a sandwich immunoassay for quantitative detection of total aggregate TDP-43 in cellular lysates using Alpha Technology.
| Feature | Specification |
|---|---|
| Application | 細胞シグナル伝達 |
| Sample Volume | 10 µL |
The AlphaLISA™ SureFire® Ultra™ Human and Mouse Total Aggregate TDP-43 assay is a sandwich immunoassay for quantitative detection of total aggregate TDP-43 in cellular lysates using Alpha Technology.
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TAR DNA binding protein 43 (TDP-43) is a DNA and RNA-binding protein which plays a crucial role in RNA metabolism. Mainly located in the nucleus, TDP-43 shuttles between the nucleus and the cytoplasm. In pathological conditions, including stress or mutation, TDP-43 is mislocalized in the cytoplasm, which can be cleaved in C terminal fragments, thus leading to an accumulation of ubiquitinated and hyperphosphorylated toxic aggregates and then insoluble inclusion bodies. Aggregated TDP-43 has been identified as a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD), and more widely in several neurodegenerative diseases such as Alzheimer’s disease.
The AlphaLISA SureFire Human and Mouse Total Aggregate TDP-43 Detection Kit is a sandwich immunoassay for the quantitative detection of total aggregate TDP-43 in cellular lysates, using Alpha Technology.
The AlphaLISA SureFire Ultra dimer/aggregate assay measures the levels of a given protein in a dimeric, multimeric or any other aggregated form in a biological sample (e.g. cell lysate).
The assay uses a single antibody clone tagged with both CaptSure™ peptide and biotin, that recognizes exposed epitopes on the multivalent 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 antibody labeled with a CaptSure tag. Donor Beads are coated with streptavidin to capture the same specific antibody, which is biotinylated. In the presence of the multivalent 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 for the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of multimeric 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 dimer or aggregate AlphaLISA SureFire Ultra detection reagents. This protocol enables cell viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
Detection of the dimer or aggregate 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 robust AlphaLISA SureFire Ultra quality.
| Application |
Cell Signaling
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
AlphaLISA SureFire Ultra
|
| Detection Modality |
Alpha
|
| Lysis Buffer Compatibility |
Lysis Buffer C
|
| Molecular Modification |
Total
|
| Product Group |
Kit
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
TDP-43
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Neuroscience
|
| Unit Size |
500 assay points
|
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The measurement of protein phosphorylation is a useful tool for measuring the modulation of receptor activation by both antibodies...
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