Revvity, 64YESTPEG, HTRF Human Total YES Detection Kit, 500 assay points

Overview
YES,also known as YES1 which belongs to the Src family of kinases, is a proto-oncogene and a non-receptor tyrosine kinase located at the plasma membrane. Once activated, YES phosphorylates various downstream effectors including LATS1/2, YAP, FAK, Caveolin-1, and p130CAS, as well as Crk. The kinase contributes to a wide range of cellular processes, including cell proliferation, survival, cytoskeleton rearrangements, migration, and adhesion. Dysregulation of YES can contribute to pathological conditions, especially in the context of cancer.
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 YES assay principle

The Total YES assay quantifies the expression level of YES in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total YES 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 YES in a cell extract, these conjugates bring the donor fluorophore into close proximity with the acceptor, generating a FRET signal. The signal 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 YES 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 YES detection reagents. This protocol allows for the cells' viability and confluence to be monitored.

 
Total YES one-plate assay protocol

Detection of Total YES 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
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-YES
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Inflammation Oncology
Unit Size-500 assay points