Revvity, 64ZATPEH, HTRF Human Total ZAP70 Detection Kit, 10,000 Assay Points

Overview
The Total ZAP-70 cellular assay is designed to monitor the expression level of ZAP-70 proteins, both phosphorylated and unphosphorylated. It is compatible with our phospho-ZAP-70 kit and enables the analysis of phosphorylated and total proteins from a single sample for a better readout of T-cell activation. ZAP-70 is a cytoplasmic protein tyrosine kinase expressed in T and NK cells, and plays a critical role in the events involved in initiating T-cell responses by the antigen receptor.
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-ZAP-70 assay principle
The Total-ZAP-70 assay quantifies the expression level of ZAP-70 in a cell lysate. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Total-ZAP-70 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 ZAP-70 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-ZAP-70 2-plate assay protocol
The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Total-ZAP-70 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

Total-ZAP-70 1-plate assay protocol
Detection of total ZAP70 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 3 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-Oncology & Inflammation
Unit Size-10,000 assay points