Revvity, 64WEE1TPEH, HTRF Human and Mouse Total Wee1 Detection Kit, 10,000 Assay Points

Overview
Wee1 is a serine/threonine kinase that plays an important role in regulating cell cylce. In response to DNA damage, Wee1 activity is upregulated to promote cell cycle arrest. Wee1 regulates the G2/M cell cycle checkpoint by phosphorylating and inactivating Cdk1. In the S phase, Wee1 phosphorylate Cdk2 preventing cells from entering the next cell cycle phase until DNA replication or repair is complete. Wee1 is overexpressed in many cancers, and has been correlated with poor disease-free survival for cancer patients. The inhibition of Wee1 is thus an active research area, especially since the emergence of new approaches involving PROTACs (PROteolysis TArgeting Chimeras).
How it works
Total Wee1 assay principle
The Total WEE1 assay quantifies the expression level of WEE1 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total WEE1 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 WEE1 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 Wee1 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 WEE1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

Total Wee1 one-plate assay protocol
Detection of Total WEE1 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
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Unit Size-10,000 assay points