Revvity, 64WDR5TPEG, HTRF Human and Mouse Total WDR5 Detection Kit, 500 assay points

Overview
WD repeat-containing protein 5 (WDR5) is a member of the WD40 protein family and plays a key role in various biological processes. It functions as a core component of the mixed lineage leukemia (MLL) histone methyltransferase complexes. WDR5 is essential for the methylation of histone H3 at lysine 4 (H3K4) on chromatin, a reaction catalyzed by the MLL1 complex, and is involved in MLL1-mediated regulation of gene transcription. Overexpression of WDR5 has been linked to a wide range of cancers, making it a prominent target for anti-cancer drug discovery efforts, including the development of PROTAC degraders.
How it works
Total WDR5 assay principle
The Total WDR5 assay measures WDR5 levels in cells. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The assay uses 2 antibodies, one labeled with a donor fluorophore and the other with an acceptor. Both antibodies are highly specific for a distinct epitope on the protein.
In the presence of WDR5 this enables an immune-complex formation involving both labeled antibodies, and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of total protein present in the sample, and provides a means of assessing the protein's phosphorylation state under a no-wash assay format.

 

Total WDR5 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 WDR5 HTRF detection reagents. This protocol allows for the cells' viability and confluence to be monitored.

Total WDR5 one-plate assay protocol
Detection of Total WDR5 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 miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 6
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-WDR5
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Oncology
Unit Size-500 assay points