Revvity, 64CDK7TPEH, HTRF Human Total CDK7 Detection Kit, 10,000 Assay Points

Overview
The Total CDK7 cellular assay monitors CDK7 protein levels.
Cyclin-dependent kinase 7 (CDK7) has dual functions in cells: CDK-activating kinase (CAK) that regulates cell cycle progression by phosphorylation of CDK1, CDK2, and CDK4/6, and is also a component of transcription factor II H (TFIIH) that regulates transcriptional initiation through phosphorylation of the RNA polymerase II C-terminal domain (CTD).
Therefore, inhibition of CDK7 has good potential for restoring a controlled cell cycle through the regulation of multiple cell cycle checkpoints, and has emerged as a new potential cancer therapy through cancer specific transcriptional regulation.
How it works
Total CDK7 assay principle
The Total CDK7 assay quantifies the expression level of CDK7 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total CDK7 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 CDK7 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 CDK7 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 CDK7 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total CDK7 one-plate assay protocol
Detection of Total CDK7 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 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-10,000 assay points