Revvity, 64CK1ATPEG, HTRF Human and Mouse Total CK1a Detection Kit, 500 Assay Points

Overview
Casein kinase 1 (CK1) is a Serine/Threonine kinase that is ubiquitously expressed in eukaryotic organisms. Among its seven isoforms, CK1 alpha is involved in a wide range of cellular processes, including membrane trafficking, cell cycle progression, chromosome segregation, apoptosis, autophagy, cell metabolism, differentiation, circadian rhythm, the immune response, neurodegeneration, and cancer.
CELMoDs (Cereblon E3 Ligase Modulating Drugs), derived from IMIDs (immunomodulatory imide drugs), represent a promising therapeutic approach in cancer treatment, particularly in hematological malignancies, due to their mechanism as molecular glues. Notably, some CELMoDs, such as Iberdomide and CC-92480, specifically target CK1A by promoting its ubiquitination and degradation, thereby inhibiting cancer cell growth and proliferation.
How it works
Total CK1A assay principle
The Total CK1A assay quantifies the expression level of CK1A in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total CK1A 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 CK1A 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 CK1A 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 HTRF Total CK1A detection reagents. This protocol permits the cells' viability and confluence to be monitored.

Total CK1A one-plate assay protocol
Detection of Total CK1A 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 allows for miniaturization while maintaining robust HTRF quality. Application-Protein Quantification
Brand-HTRF
Buffer/Solvent-Lysis Buffer 1
Detection Modality-HTRF
Host Species-Human Mouse
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-CK1a
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Inflammation Oncology
Unit Size-500 assay points