Cellular Analysis
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  • Reporter Systems

    Studies of protein expression, interactions, and structure, often use reporter systems to introduce and select for gene targets in cells.  Reporter genes confer drug resistance, bioluminescence or fluorescence properties into the cells they are introduced. Typical reporter studies link reporter genes directly to a promoter region of interest, the function of which can be monitored  by the reporter activity. Protein fusion tagging is used to detect subcellular localization, degradation, protein-protein interactions, etc. Typical fusion tags are fluorescent proteins (e.g eGFP) or small protein epitopes (e.g FLAG, Myc HA) which can be detected by fluorescence FACS or western blots. New generations of reporter gene systems expand the range of applications  and enhance experimental possibilities.

    • Luciferase reporter genes with the enhanced properties of greater stability, robust secretory signal, smaller size, high sensitivity and no dependence on ATP are applicable for reporter gene studies, siRNA potency screening, live cell monitoring and BRET applications in vivo.
    • Enhanced GFP and GFP color derivatives are useful for imaging mammalian cells, single fluorescent protein-based biosensors, FRET and producing transgenic model organisms.  However GFP can be toxic to cells at high expression levels.
    • Multimodality fusion reporter technologies employ vectors containing multiple reporter genes.  These systems enable optimal analysis by multiple methods, such as fluorescence for single cell imaging, bioluminescence for small animal imaging, or short-lived radioactive tracers in living subjects for PET.
    • Split reporter systems enable protein-protein interaction assays by cloning target genes into separate vectors, each linked to complementary portions of a reporter, such as luciferases or GFP.

    Activity of Gaussia (GLuc) and Cypridina (CLuc) Luciferases Expressed in CHO Cells.

    Activity was determined from the conditional media or cell lysate.

    Extreme Sensitivity of the Gaussia Luciferase Reporter.

    HeLa cells were transfected with 1 µg of pCMV-GLuc or pCMV-Renilla, and activity was measured from the cell supernatant (S) or cell lysate (L). Note that although Gaussia Luciferase is a secreted reporter, there is sufficient measurable activity inside the cell. Renilla is only expressed intracellularly.

    Featured Products

    Reporter Systems includes these areas of focus:

    Cypridina Luciferase
    Gaussia Luciferase

    Transfection Reagents Selection Chart (Discontinued)

    NEB offers a broad selection of high efficiency transfection reagents suitable for transfecting DNA, siRNA and protein.

    NEB vs. Other Commercially Available Reporter Systems

    Features Gaussia Cypridina Renilla Firefly Metridia
    Transcriptional
    Reporter
    Extreme Sensitivity      
    Secreted    
    Stable      
    Does not require and is
    not affected by ATP
     

    Comparison of the features of commercially available reporter systems highlights the advantages of using Gaussia or Cypridina Luciferase from NEB.

    Reporter System Selection Chart

    NEB offer a variety of products that utilize secreted Gaussia Luciferase (GLuc) and Cypridina Luciferase (CLuc) in reporter assays.