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The AlphaLISA™ SureFire® Ultra™ Human and Mouse Total SQSTM1 p62 assay is a sandwich immunoassay for quantitative detection of total SQSTM1 p62 in cellular lysates using Alpha Technology.
| Feature | Specification |
|---|---|
| Application | 細胞シグナル伝達 |
| Protocol Time | 2h at RT |
| Sample Volume | 10 µL |
The AlphaLISA™ SureFire® Ultra™ Human and Mouse Total SQSTM1 p62 assay is a sandwich immunoassay for quantitative detection of total SQSTM1 p62 in cellular lysates using Alpha Technology.
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Sequestome-1 or ubiquitin binding protein p62 (SQSTM1 p62) is an autophagosome cargo protein that targets other proteins that bind to it for selective autophagy. SQSTM1 p62 is an adaptor protein which is initially phosphorylated by ULK1 on Serine 407, then on Serine 403 by casein kinase 2 or TBK1. The phosphorylation on Ser403 increases its affinity for ubiquitin chains, thus enabling SQSTM1 p62 to bind to ubiquitinated cargo proteins. Ubiquitinated proteins or ubiquitin coated mitochondria associated with SQSTM1 p62 proteins are taken away in phagosomes, whose content is cleared out after lysosomal fusion. SQSTM1 p62 is implicated in Alzheimer’s disease, Huntington’s disease, NASH, and numerous types of cancer.
The AlphaLISA SureFire Ultra Human and Mouse Total SQSTM1 p62 Detection Kit is a sandwich immunoassay for the quantitative detection of total SQSTM1 p62 in cellular lysates, using Alpha Technology.
The Total-AlphaLISA SureFire Ultra assay measures the expression level of a target protein in a biological sample (e.g. cell lysate).
The Total-AlphaLISA SureFire Ultra assay uses two antibodies which recognize two different distal epitopes on the 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 one of the detection antibodies, which is biotinylated. In the presence of target 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 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 enables cell 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 robust AlphaLISA SureFire Ultra quality.
| Application |
Cell Signaling
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
AlphaLISA SureFire Ultra
|
| Cellular or Signaling Pathway |
Mitophagy
|
| Detection Modality |
Alpha
|
| Lysis Buffer Compatibility |
Lysis Buffer
|
| Molecular Modification |
Total
|
| Product Group |
Kit
|
| Protocol Time |
2h at RT
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
p62 (SQSTM1)
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Neuroscience
Oncology
|
| Unit Size |
50,000 assay points
|
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