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| Feature | Specification |
|---|---|
| Application | 細胞シグナル伝達 |
| Sample Volume | 10 µL |
Cyclin D3 is an atypical D-type cyclin that is primarily expressed in differentiated tissues. Overall, D-type cyclins (D1, D2, and D3) are involved in the regulation of transition from G1 to S during the cell cycle. Cyclin D activity is regulated through its mitogen-activated transcription, binding to CDK4/6, phosphorylation, ubiquitylation, nuclear export, and degradation. Cyclin D3 plays a key role in differentiation and maturation in lymphoid and musculoskeletal tissues. Increased expression of Cyclin D3 is associated with cells developing resistance to the CDK4 inhibitor Palbociclib, suggesting it as an effective therapeutic target.
The AlphaLISA SureFire Ultra Human Total Cyclin D3 Detection Kit is a sandwich immunoassay for the quantitative detection of total Cyclin D3 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 |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
Cyclin D3
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Oncology
|
| Unit Size |
10,000 assay points
|
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