Revvity, 64CDK12TPEH, HTRF Human Total CDK12 Detection Kit, 10,000 Assay Points

Overview
The Total CDK12 cellular assay monitors CDK12 protein levels.
CDK12 is a transcription-associated CDK. It complexes with cyclin K to regulate gene transcription elongation via the phosphorylation of RNA polymerase II (RNAP II), and also regulates translation. Moreover, CDK12 plays a role in RNA splicing, cell cycle progression, cell proliferation, DNA damage response (DDR), and the maintenance of genomic stability. Since the mutation or amplification of CDK12 is closely related with tumorigenesis, CDK12 has become an attractive therapeutic target for cancer treatments.
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 CDK12 assay principle
The Total CDK12 assay quantifies the expression level of CDK12 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total CDK12 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 CDK12 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 CDK12 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 CDK12 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total CDK12 one-plate assay protocol
Detection of Total CDK12 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 2
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-10,000 assay points