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The AlphaLISA™ SureFire® Ultra™ Human Total Cyclin D1 assay is a sandwich immunoassay for quantitative detection of total cyclin D1 in cellular lysates using Alpha Technology.
| Feature | Specification |
|---|---|
| Application | Cell Signaling |
| Sample Volume | 30 µL |
The AlphaLISA™ SureFire® Ultra™ Human Total Cyclin D1 assay is a sandwich immunoassay for quantitative detection of total cyclin D1 in cellular lysates using Alpha Technology.
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Cyclin D1 is encoded by CCND1 and is a member of the highly conserved cyclin family, which includes Cyclin D2 and Cyclin D3. Cyclin family members are important regulators of cell cycle progression and act as mediatiors of growth factor dependent G1 to S phase progression by playing the role of allosteric regulators of CDK4/6. Cyclin D1 is an oncogene and is found to be overexpressed in many human cancers. Abnormal levels of Cyclin D1 result in dysregulated CDK4/6 activity and leads to tumorigenesis. Cyclin immunoassays can quantify cyclin.
The AlphaLISA SureFire Human Total Cyclin D1 Detection Kit is a sandwich immunoassay for the quantitative detection of total Cyclin D1 in cellular lysates, using Alpha Technology.
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 |
Cyclin D1
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
Mouse
|
| Technology |
Alpha
|
| Therapeutic Area |
Oncology
|
| Unit Size |
100 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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