Revvity, 64TIE2TPEG, HTRF Human Total Tie2 Detection Kit, 500 assay points

Overview
The Total Tie2 cellular assay kit is designed to monitor the expression level of Tie2, whether phosphorylated or unphosphorylated. It is compatible with our Phospho-Tie2 kit, and enables the analysis of phosphorylated and total proteins from a single sample for better readouts of the angiopoietin/Tie2 signaling pathway.
Tie2 is a tyrosine-protein kinase found in endothelial cells, functioning as a cell-surface receptor for angiopoietins. It plays a key role in regulating angiogenesis, cell migration, survival, adhesion, and proliferation, with its primary function being to maintain vascular stability and quiescence.
How it works
Total Tie2 assay principle
The Total Tie2 assay quantifies the expression level of Tie2 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total Tie2 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 Tie2 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 Tie2 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 Tie2 detection reagents. This protocol allows for the cells' viability and confluence to be monitored.

Total Tie2 one-plate assay protocol
Detection of Total Tie2 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 1
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-Tie2
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology
Unit Size-500 assay points