Revvity, 64METTPEG, HTRF Human Total MET Detection Kit, 500 Assay Points

Overview
c-MET, also called tyrosine-protein kinase MET or hepatocyte growth factor receptor (HGFR), is a human MET gene-coded protein with tyrosine kinase activity. It acts as a tyrosine kinase receptor, with functions related to wound healing, organogenesis, and embryonic development.
MET is expressed in epithelial cells and is triggered by hepatocyte factors HGF and HSF, with ramifications into various pathways like RAS-MAPK, AKT, and STAT3 signaling.
Abnormal activation of MET is linked to cancer progression, with effects on tumor growth, angiogenesis, and tumor cell migration & invasion. These relationships between MET and tumor cells are found across multiple cancer types (liver, brain, breast, kidney, etc) and are suspected to originate from a hijacking of MET and its ligand expression regulation, which leads to their overexpression and autocrine activation of MET.
How it works
Total MET assay principle
The HTRF Total MET assay quantifies the expression level of MET in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The Total MET assay uses two labeled antibodies, one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of MET 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 MET two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a low volume detection plate (either HTRF 384-lv or 96-lv plate) before the addition of HTRF Total MET detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total MET one-plate assay protocol
Detection of Total MET 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 Lysis Buffer 2
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
Unit Size-500 assay points