Revvity, 64CDK1TPEG, HTRF Human Total CDK1 Detection Kit, 500 Assay Points

Overview
CDK1, also known as Cell Division Cycle 2 (Cdc2), is a serine/threonine kinase that is ubiquitously expressed in eukaryotes. CDK1 is a catalytic subunit of a protein kinase called the M-phase promoting factor that induces entry into mitosis. CDK1 promotes G2-M transition, and regulates G1 progress and G1-S transition via association with multiple interphase cyclins (Cyclin A, Cyclin B). Dysregulated activation of CDK1, leading to uncontrolled cell division, is a hallmark of cancers. The inhibition of CDK1 is thus an active research area, especially since the emergence of new approaches involving PROTACs (PROteolysis TArgeting Chimeras).
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
Total CDK1 assay principle
The Total CDK1 assay quantifies the expression level of CDK1 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total CDK1 assay uses two labeled antibodies, one coupled to a donor fluorophore and the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In the presence of CDK1 in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein's expression under a no-wash assay format.

 

Total CDK1 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Total CDK1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total CDK1 one-plate assay protocol
Detection of Total CDK1 with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-500 assay points