Nuclear Protein Product Solutions

Cell-Based Assays to Evaluate Activation and Translocation of Nuclear Proteins

Nuclear proteins are key targets for drug development due to their ubiquitous roles in controlling cellular behavior and pathway expression. The translocation of many nuclear proteins into and out of the nucleus is a highly controlled mechanism that can dictate protein function. Controlling this translocation behavior using therapeutics, or ensuring that a potential drug does not cause undesired effects on translocation, are important applications for measuring nuclear translocation.


Nuclear hormone receptors (NHRs), including steroid receptors, are a critical subset of nuclear proteins that regulate gene expression in response to ligand activation and are intimately involved in endocrine pathway signaling. NHRs commonly rely on a coactivator or corepressor protein, often a steroid receptor cofactor (SRC protein), to perform their function. Enhancing or disrupting the association of these proteins can be a target for therapeutic development focused on controlling NHR-related gene expression.


Eurofins DiscoverX® provides PathHunter® nuclear translocation and protein interaction cell-based, functional assays to investigate protein translocation events and activation of nuclear proteins. Nuclear translocation assays allow for studying the trafficking of your protein of interest to and from the nucleus and analyzing the pharmacological effects of therapeutics on these events. Protein interaction assays can be used to evaluate the interaction of NHRs and their cofactors, including therapeutics that affect this protein-protein interaction.


Among the advantages of the Eurofins DiscoverX platform is that, unlike other assays, these cell-based assays do not require antibodies, fluorescent tags, or specialized equipment, and they provide an easy-to-use, high-throughput capable, and quantitative measurement of protein-protein interaction.


Advantages of Nuclear Protein Assays

  • Homogeneous, HTS-compatible Assay – An easy-to-use, sensitive, and high-throughput cell-based assay platform without the need for antibodies, fluorescent tags, or specialized equipment
  • Flexible Solutions – Create your own quantitative, functional cell-based assays to study nuclear translocation of any desired target protein
  • Diverse Therapeutic Discovery – Quantify small molecules, peptides, or antibody-based therapeutics targeting nuclear proteins

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Related Topics

Nuclear Protein Assays

Custom and Toolbox Products

  • Custom Capabilities – Custom cell lines and assay capabilities optimized to fit your requirements
  • Toolbox Products – Complete set of parental cell lines, expression and cloning vectors, kits, and retroparticles to build your own stable cell lines and cell-based assays

Nuclear Translocation Assay Targets

Keap1-NRF2 RELA TORC1 (CRTC1) TORC3 (CRTC3) β-Catenin (Wnt-FzGSK3β)

NHR Nuclear Translocation and Protein Interaction Targets


PathHunter nuclear protein assays are based on the proprietary Enzyme Fragment Complementation (EFC) technology to measure the movement of proteins into the nucleus or the protein-protein interactions in the nucleus.

Nuclear Translocation Assay Principles

Nuclear translocation assays allow for the identification, screening, lead optimization, and even safety testing of therapeutics. These assays monitor the activation or inhibition of a particular signaling pathway coupled with the movement of proteins from the cytosolic compartment to the nuclear compartment. In general, this approach enables you to create a highly specific, non-transcriptional assay for almost any protein that is known to translocate to the nucleus, such as FOXO3, NRF2, XBP1, p53, and many others. Learn about several translocation assays on the nuclear proteins and signaling pathways pages.

NHR Nuclear Translocation Assay Principle

NHR Nuclear Translocation Assay Principle

NHR translocation assays detect the binding of small molecules that trigger NHR activation and translocation into the nucleus. The cells have been engineered to express the ED fragment of the β-gal enzyme fused to the NHR (that is located in the cytoplasm) and the EA fragment localized in the nucleus. Ligand binding to the NHR induces the receptor translocation from the cytoplasm to the nucleus, forcing the complementation of the ED and EA fragments and the formation of an active β-gal enzyme. The enzyme then hydrolyzes a substrate to generate a chemiluminescent signal that can be easily measured on any standard luminometer.

Measure Activation-Induced Nuclear Translocation of Transcription Factors

Measure Activation-Induced Nuclear Translocation of Transcription Factors

Quantitative data showing binding and ligand-induced translocation on NHRs such as androgen, glucocorticoid, liver X, mineralcorticoid, and progesterone receptors. Monitoring Glucocorticoid Receptor Nuclear Translocation. A. Immunofluorescence analysis of dexamethasone induced Glucocorticoid Receptor translocation into the nucleus using PathHunter CHO-K1 Glucocorticoid Receptor Nuclear Translocation cells. B. Translocation EFC-based assay (Cat. No. 93-0002C2) of the same cells stimulated with dexamethasone for 3 hours and results shown an EC50 of 8.2 nM and excellent signal-to-background of 15.9. PathHunter NHR translocation assays employ full length NHRs providing more biologically relevant data, and shorter assay times allowing for fewer off-target effects.