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AlphaLISA SureFire Ultra Human Total Cyclin B1 Detection Kit, 500 Assay Points

The AlphaLISA™ SureFire® Ultra™ Human Total Cyclin B1 assay is a sandwich immunoassay for quantitative detection of total Cyclin B1 in cellular lysates using Alpha Technology.

Feature Specification
Application Cell Signaling
Sample Volume 10 µL

The AlphaLISA™ SureFire® Ultra™ Human Total Cyclin B1 assay is a sandwich immunoassay for quantitative detection of total Cyclin B1 in cellular lysates using Alpha Technology.

Product variants
Unit Size: 500 assay points
Part #:
ALSU-TCYCB1-A500
List price
USD 2,392.92
Your online price:
Unit Size: 10,000 assay points
Part #:
ALSU-TCYCB1-A10K
List price
USD 14,400.00
Your online price:
Unit Size: 50,000 assay points
Part #:
ALSU-TCYCB1-A50K
List price
USD 46,000.00
Your online price:
Unit Size: 100 assay points
Part #:
ALSU-TCYCB1-A-HV
List price
USD 625.00
Your online price:
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

Overview

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.

Formats
  • The HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 μL reaction volume.
  • The 500-point kit contains enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.
  • The 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.
  • The 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.
AlphaLISA SureFire Ultra kits are compatible with
  • Cell and tissue lysates
  • Antibody modulators
  • Biotherapeutic antibodies
Alpha SureFire kits can be used for
  • Cellular kinase assays
  • Receptor activation studies
  • Screening

How it works

Total-AlphaLISA SureFire Ultra assay principle

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.

assay-principle-Total-AlphaLISA-Surefire-Ultra.jpg

 

Total-AlphaLISA SureFire Ultra two-plate assay protocol

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.

2 plates assay protocol AlphaLISA Surefire Ultra Total assay

Total-AlphaLISA SureFire Ultra one-plate assay protocol

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.

1-plate-assay-protocol-AlphaLISA-Surefire-Ultra-Total-assay

Specifications

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

Video gallery

Resources

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Technical Note Icon
Technical Note
Detecting changes in phosphorylation events during the cell cycle

Lamin A/C, a crucial nuclear protein, undergoes phosphorylation at mitotic sites, controlled by the CDK1-Cyclin B1 complex...

Technical Note Icon
Technical Note
PROTAC degraders targeting CDK4/CDK6 for the study of cell cycle regulation in oncology

The cell cycle is tightly regulated by key proteins like Cyclin D1, which forms complexes to initiate progression through phases...

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