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.

   Check our lysis buffer compatibility

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.

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
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Phospho-total protein blocking reagent - 2 ml64KB1AAC
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Phospho-total protein blocking reagent - 6 ml64KB1AAD
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Phospho-total protein lysis buffer #1 - 130 mL64KL1FDF
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Literature