Epigenetic Proteins Product Solutions

Epigenetics involves the study of heritable or acquired changes in gene expression caused by non-genetic mechanisms without alterations in gene structure or sequence. Basic epigenetic mechanisms such as histone modifications, DNA methylation, and chromatin remodeling are essential in the regulation of many physiological processes. Alterations to the mechanisms, either by modifications to DNA and histone components of nucleosomes, or expression of noncoding RNAs (ncRNAs) can have implications in a number of cancers, immunological, and neurodegenerative conditions.

 

Eurofins DiscoverX offers epigenetics InCELL Hunter™ cell-based assays for studying the modulation of epigenetic proteins and identifying small molecule inhibitors and modifiers of DNA methyltransferases and bromodomains. Utilize InCELL Hunter cell-based compound-target engagement assays to confirm small molecule compound cell entry and epigenetic target binding, screen and rank inhibitors, validate hits identified in biochemical assays, and measure cellular EC50 values all in the native cellular environment.

 

Read App Note

 

Product Highlights

  • Biologically Relevant - Obtain quantitative cellular compound entry and epigenetic target binding data for more confident drug discovery decisions
  • Easy-to-Use - Simple, homogeneous protocol to easily measure compound binding to your epigenetic target with a chemiluminescent readout
  • High-Throughput Amenable - Easily automate InCELL assays to screen multiple compounds accurately
  • Target Specificity & Selectivity - Ability to screen and differentiate multiple target-selective stereoisomers of BRD4 and determine target specificity

Key Resources

Application Note: Automating a Cell-Based Target-Compound Assay for Methyltransferase and Bromodomain.

Scientific Posters & Presentations

Related Topic: InCELL Target Engagement Assays

Products

  • InCELL Hunter Epigenetic Cell Lines - Stable cell lines confirmed for past 10 passages and rigorously and continuously validated for accurate pharmacology through Eurofins Discovery services with billions of data points screened.
  • InCELL Hunter eXpress Kits - Complete assay-ready kits with specific cell line, optimized reagents, and plates for quick Enzyme Fragment Complementation (EFC) based chemiluminescent detection.
  • InCELL Pulse Starter Kit - Create your own cell-based, target engagement assays with reagents and protocol for the design and development of InCELL Pulse assays for the measurement of inhibitor cellular potency against user-defined epigenetic targets of interest.
  • InCELL Detection Kit - Measure compound binding to a specific protein target in InCELL Hunter and InCELL Pulse cell lines

Custom Products

  • Custom Capabilities - Custom cell lines, assays, and protein development capabilities optimized to your requirements.

InCELL Hunter Assay Principle

InCELL Pulse Assay Principle

InCELL Hunter stabilization assays are based on the industry validated Enzyme Fragment Complementation (EFC) technology and built on the principle of protein stabilization related to the protein turnover. Cells expressing a protein of interest is fused to the small EFC enzyme donor of β-galactosidase (β-gal) called ePL (enhanced ProLabel®). Upon addition of a compound that binds the target, protein levels are stabilized or altered in the cell, and this change can be monitored by measuring target protein abundance using chemiluminescent detection. Upon addition of the large EFC enzyme acceptor (EA) fragment and chemiluminescent substrate, EA naturally complements with the ePL tag on the target protein to create an active β-gal enzyme. The resulting active enzyme hydrolyzes the substrate to generate a chemiluminescent signal. The greater the signal, the greater the presence of compound-target engagement in the cell.

 

InCELL Pulse Assay Principle

InCELL Pulse Assay Principle

InCELL Pulse™ compound-target engagement assays are based on the EFC technology incorporating compound binding detection based on protein thermal stability. Cells expressing a protein of interest is fused to the small EFC β-gal enzyme donor ePL. These cells are treated with test compound and then subjected to elevated temperatures during a pulse denaturation step. The larger EFC enzyme donor (EA) of β-gal is then added with a chemiluminescnet substrate and EFC complementation. Compound binding protects the target protein from thermal denaturation, which enhances complementation between added EA and ePL components and increases the chemiluminescent signal measured using the EFC-based detection system. In the absence of compound binding, the target protein forms denatured aggregates that poorly complement with EA, which results in a low chemiluminescent signal.