Revvity, 63ADK015PEG, HTRF Human and Mouse Phospho-ATF2 (Thr71) Detection Kit, 500 Assay Points

Overview
ATF2 (Activating transcription factor 2) is a transcription factor activated by phosphorylation on threonin residues by JNK/SAPK. The phospho-ATF2 (Thr71) kit enables the quantitative detection of transcription factor ATF2 modulation phosphorylated at Threonine-71. ATF2 is also involved in PI3K/AKT, TNF, and estrogen signaling along with insulin secretion, thyroid hormone release, substance dependence, viral carcinogenesis and infectious diseases. The simple add-and-read protocol features no wash steps for a faster analysis in biological applications like oncology, diabetes or inflammation.
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
Phospho-ATF2 (Thr71) kit assay principle
The Phospho-ATF2 (Thr71) assay measures ATF2 when phosphorylated at Thr71. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-ATF2 (Thr71) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation 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 phosphorylated protein present in the sample, and provides a means of assessing the protein's phosphorylation state under a no-wash assay format.

 

Phospho-ATF2 kit 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 Phospho-ATF2 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Phospho-ATF2 kit 1-plate assay protocol
Detection of Phosphorylated ATF2 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-Phosphorylation
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 Metabolism/Diabetes
Unit Size-500 assay points