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| Feature | Specification |
|---|---|
| Application | Cell Signaling |
| Sample Volume | 10 µL |
G2/mitotic-specific cyclin-B1 (Cyclin B1) is a key component in the control of cell cycle progression from G2 to M phase and has been implicated in the tumorigenesis and the development of malignancy. Under DNA damage conditions, Cyclin B1 becomes upregulated, presumably already in the DNA replication phase (S-phase). This upregulation is controlled by SOG1, which in turn is activated by the checkpoint kinases ATM and ATR, which sense DNA damage. Cyclin B1/CDK1 regulate mitochondrial bioenergetics for cell cycle progression and tumor response to therapy. Cyclin B1 is associated with breast, colorectal, ovarian, lung, and pancreatic cancer.
The AlphaLISA SureFire Human Total Cyclin B1 Detection Kit is a sandwich immunoassay for the quantitative detection of total Cyclin B1 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 |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
Cyclin B1
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Oncology
|
| Unit Size |
500 assay points
|
Are you looking for resources, click on the resource type to explore further.
Lamin A/C, a crucial nuclear protein, undergoes phosphorylation at mitotic sites, controlled by the CDK1-Cyclin B1 complex...
The cell cycle is tightly regulated by key proteins like Cyclin D1, which forms complexes to initiate progression through phases...
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