Revvity, 63ADK096PEG, HTRF Human and Mouse Total STAT1 Detection Kit, 500 Assay Points

Overview
The total STAT1 assay measures endogeneous levels of STAT1 in cell lysates. STAT1 acts as an important transcriptional activator and the assay serves as a readout for JAK inhibitors in oncology, virology, and inflammation. The assay can be used to normalize the amount of phosphorylated STAT1 over total STAT1.
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-STAT1 assay principle
The Total-STAT1 assay quantifies the expression level of STAT1 in a cell lysate. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Total-STAT1 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 STAT1 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 proteins expression under a no-wash assay format.

 

Total-STAT1 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-STAT1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-STAT1 1-plate assay protocol
Detection of total STAT1 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
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Infectious Diseases Neuroscience Oncology & Inflammation
Unit Size-500 assay points