Phospho-ATF2 (Thr71) Cellular Assay Kit

Homogeneous cell-based assay for phospho-ATF2 as a readout of JNK1 or p38MAPK

Cisbio's cell-based homogeneous HTRF® phospho-ATF2 assay (Thr71) enables both SAPK/JNK and p38 MAPK pathway readout, giving quantitative detection of the modulation of the transcription factor ATF2, phosphorylated on Threonine 71. ATF2 is also involved in PI3K/AKT, TNF and estrogen signaling, and amongst others ATF2 plays a role in insulin secretion, thyroid hormone release, substance dependence, viral carcinogenesis and infectious diseases. The kit is suitable for analysing pharmacological questions in cellular models. The simple add-and-read protocol, with no wash steps for a faster analysis, facilitates a wide variety of biological applications from basic reasearch to high throughput screening in oncology, diabetes, inflammation, cardiovascular, virology and CNS areas.

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Assay principle

HTRF® - the homogeneous cell-based sandwich immunoassay

Cisbio's phospho-ATF2 assay is based on a TR-FRET sandwich immunoassay format comprising two specific anti-ATF2 antibodies, one labeled with a cryptate as donor and the other labeled with d2 as acceptor. The phospho-ATF2 antibodies bind the phosphorylated residue and the proximity of donor and acceptor will then lead to a fluorescent TR-FRET signal. The signal intensity is proportional to the substrate phosphorylation. The protocol is optimized for a 384-well plate format, but can easily be further miniaturized or upscaled. Only low sample volumes are needed. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. No washing is needed at any step.

The phospho-ATF2 assay kit can be run with frozen cell lysates or fresh cells in culture. After cell lysis, phospho-ATF2 can be quantitatively detected using the HTRF phospho-ATF2 kit reagents and most TR-FRET multimode plate readers.

Two-plate assay protocol

For added flexibility, the assay can be run under a two-plate assay protocol, where cells are plated and treated in a 96-well culture plate. For detection of phosphorylated ATF2, lysates are subsequently transferred to a 384 small volume assay plate where the HTRF reagents are added. This also enables the monitoring of cell viability and confluence in an appropriate cell culture plate.

What to expect at the bench

This video covers the principles of phospho-HTRF assays and gives guidelines for performing them, using our phospho-ERK assay as an example.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
ATF2 phospho-T71 kit - 500 tests63ADK015PEG
ATF2 phospho-T71 kit - 10,000 tests63ADK015PEH
ATF2 phospho-T71 kit - 50,000 tests63ADK015PEY

Companion products

DescriptionCat. noProduct insertMSDS
ATF2 phospho-T71 kit - control lysate63ADK015TDA