Multiplexed Measurement of Protein Levels, Transcriptional Activation, and Cell Viability

2 May 2017

Cellular stress responses often involve changes in the stability of a critical regulatory protein, followed by downstream modulation of specific transcriptional targets. This application note describes a multiplexed assay of protein level, transcriptional activation, and cell viability in the same well using NanoDLR™ and CellTiter Fluor™ Assays on the GloMax® Discover System. It also demonstrates how multiplexed measurements of protein abundance, gene expression, and cell viability can greatly increase the amount of information obtained from a single sample.

GloMax® Discover Microplate Reader

Promega Corp.

  This ready-to-use multimode detection system includes: high-performance luminescence, fluorescence, UV-Visible absorbance, BRET and FRET, two-color filtered luminescence and kinetic measurement capabilities.   The GloMax® Discover System is a high-performance multimode detection instrument developed with Promega reagent chemistries to provide a simple means of detecting advanced chemistries. This instrument provides researchers superior luminescence sensitivity and dynamic range for both strong and weak bioluminescence experimental samples, as well as a seamless integration with Promega’s industry leading bioluminescent assays. GloMax® Discover also provides flexible use of filters for fluorescence intensity, BRET, FRET, filtered luminescence and UV-visible absorbance measurements. The instrument is operated by an integrated Tablet PC, which provides quick and easy navigation through the control options. Exporting your results is made seamless with a variety of options, including exporting to your local data network. The GloMax® Discover software provides the required technical elements of a 21 CFR Part 11-compliant system to be used with the appropriate laboratory workflow. GloMax® Discover System Features: Integrated with Promega Assays - Developed and optimized with Promega cell and gene reporter assays for seamless workflow; you will be up and running faster. Easy-to-Use - Simple Tablet PC touch screen navigation with full PC capabilities and a state-of-the-art graphical user interface. Superior Performance - Broader dynamic range, better sensitivity and lower well-to-well cross talk for more usable data from your experiment. Cost-Effective - Spend less time optimizing. Connected to Your Workflow - Use as a standalone instrument or integrate into your high-throughput automated workflow; export data to your laboratory network; enable 21 CFR, Part 11 electronic signature compliance. Applications: Integration with Promega reagent chemistries for detection of fluorescence intensity, BRET, FRET, filtered luminescence and UV-visible absorbance assays. Integration into low- and medium-throughput automation workflows. Optimization with Promega cell and gene reporter assays.

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ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell AnalysisThe analysis of cells allows researchers to understand the factors which contribute to cell health and function. These influencing processes can then be predicted and altered, leading to the development of medication and disease treatments.Assay DevelopmentThe process of proving an assay to be sensitive with respect to the target is known as assay development. The assay should be able to characterize novel compounds and measure the potency of these compounds against a validated biological target.TranscriptomicsHypoxiaGene RegulationProtein BiologyMultiplexingMultiplexing refers to the ability to measure multiple targets or analytes simultaneously in a single experiment. This technique is valuable for high-throughput screening, diagnostics, and complex assays, as it increases efficiency and data quality. Browse our peer-reviewed product directory to find the best multiplexing products, compare tools, check reviews, and get pricing directly from manufacturers.