View All
| Feature | Specification |
|---|---|
| Application | Cell Signaling |
| Sample Volume | 30 µL |
TFE3, or transcription factor E3, is a member of the basic helix-loop-helix leucine zipper MiT transcription factory family. MiT/TFE transcription factors play essential roles in the regulation of autophagy and lysosomal biogenesis. Phosphorylation is the primary regulator of MiT/TFE. The phosphorylated protein remains in the cytoplasm and, following dephosphorylation, translocates to the nucleus, where it induces autophagy and lysosomal biogenesis by promoting the expression of hundreds of genes. Several signaling molecules involved in the mTORC1, PKC, and AKT pathways can directly influence the transcription factor-mediated lysosome-to-nucleus signaling. TFE3 is associated with Parkinson’s disease, Alzheimer’s diseases, and various types of cancer.
The AlphaLISA SureFire Ultra Human and Mouse Total TFE3 Detection Kit is a sandwich immunoassay for the quantitative detection of total TFE3 in cellular lysates, using Alpha Technology.
Formats:
AlphaLISA SureFire Ultra kits are compatible with:
Alpha SureFire kits can be used for:
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 |
30 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
TFE3
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Neuroscience
Oncology
|
| Unit Size |
100 assay points
|
Are you looking for resources, click on the resource type to explore further.
The definitive guide for setting up a successful AlphaLISA SureFire Ultra assay
Several biological processes are regulated by...
Discover Alpha SureFire® Ultra™ assays, the no-wash cellular kinase assays leveraging Revvity's exclusive bead-based technology...
This document includes detailed tables listing HTRF™, AlphaLISA™ SureFire® Ultra™, and Alpha SureFire® Ultra™ Multiplex assays...
Loading...
We are here to answer your questions.