Phospho-MNK (Thr197/202) Cellular Assay Kit

HTRF® assay for studying the MNK/EIF4e translational pathway in cancer studies

MNK1 is a kinase involved in the MAPK, HIF1 and insulin pathways. Upon phosphorylation by ERK, MNK1 phosphorylates CREB (MAPK) or EIF4E (HIF1 and insulin) thus playing a key role in mRNA translational regulation.

Cisbio's cell-based homogeneous HTRF® phospho-MNK assay Thr197/202 enables quantitative detection of the modulation of the CAM kinase of the MAPKKAPK family MNK1, phosphorylated on Threonine 197/202.

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Assay principle

HTRF® - the homogeneous cell-based sandwich immunoassay

Cisbio's phospho-MNK assay is based on a TR-FRET sandwich immunoassay format comprising two specific anti-MNK antibodies, one labeled with a cryptate as donor and the other with d2 as acceptor. The phospho-MNK 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-MNK assay kit can be run with frozen cell lysates or fresh cells in culture. After cell lysis, phospho-MNK can be quantitatively detected using the HTRF phospho-MNK 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 MNK, 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
MNK phospho-T197/202 kit - 500 tests63ADK022PEG
MNK phospho-T197/202 kit - 10,000 tests63ADK022PEH

Companion products

DescriptionCat. noProduct insertMSDS
MNK phospho-T197/202 kit - control lysate63ADK022TDA
Phospho-total protein blocking reagent - 2 ml64KB1AAC
Phospho-total protein blocking reagent - 6 ml64KB1AAD
Phospho-total protein lysis buffer #1 - 130 mL64KL1FDF