Revvity, 64PLCG2TPEG, HTRF Human Total PLC-γ2 Detection Kit, 500 Assay Points

Overview
This HTRF cell-based assay conveniently and accurately detects Phospholipase C gamma 2 (PLCgamma 2) protein.
PLCgamma 2 is a phosphoinositide-specific phospholipase that generates second messengers and plays a crucial role in signal transduction, especially in haematopoietic cells such as B cells, mast cells, macrophages, natural killer cells and platelets. Upon stimulation of different receptors such as B-cell antigen receptor (BCR) and Fc receptors (FcRs), PLCgamma 2 interacts with several adapter proteins and becomes phosphorylated by intracellular tyrosine kinases. Phosphorylation includes several sites, one of which is Tyr1217.
PLCgamma 2 has been linked to various pathologies such as auto-immune and inflammatory diseases, drug-resistance in cancer and neurodegeneration.
How it works
Total-PLCg2 assay principle
The Total PLCg2 assay quantifies the expression level of PLCg2 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-PLCg2 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 PLCg2 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-PLCg2 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the addition of Total-PLCg2 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-PLCg2 one-plate assay protocol
Detection of Total PLCg2 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 4 Lysis Buffer 5
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
Therapeutic Area-Neuroscience Oncology & Inflammation
Unit Size-500 assay points