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GloResponse 9XGAL4UAS-luc2P HEK293 Cell Line

by Promega
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  • Protocols
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  • Live-Cell Detection of GAL4-Mediated Signaling Events

    The GloResponse™ Luciferase Reporter Cell Lines contain optimized, state-of-the-art luciferase reporter technology integrated into a cell line. This allows the rapid development of a reporter assay based on the pathway of interest regulating the luciferase gene. Assays configured using the GloResponse™ Cell Lines are suitable for high-throughput screening, typically having greater response dynamics (fold of induction) than other assay formats and good quality as indicated by high Z´ values. GloResponse™ Cell Lines were developed to study a variety of signaling pathways. Activators of these pathways may be native to the HEK293 cell line. Activity of non-native activators can be studied after they have been introduced by transfection.

      The GloResponse™ 9XGAL4UAS-luc2P Cell Line is a clonal derivative of Human Embryonic Kidney 293 (HEK293) cells. These cells contain nine repeats of GAL4 UAS (Upstream Activator Sequence). This sequence drives transcription of the luciferase reporter gene luc2P in response to binding of a fusion protein containing the Gal4 DNA Binding Domain (e.g., the Estrogen Receptor Ligand Binding Domain in pBIND-ER? Vector [Cat.# E1390]) when activated by a ligand. This makes the cell line suitable for the study of nuclear receptors or can be used to study other types of protein:protein and protein:DNA interactions. The GAL4 DNA Binding Domain partner must be introduced to this cell line by transfection or other similar techniques.

      The GloResponse™ Cell Lines incorporate the improvements developed for the pGL4 family of reporter vectors for enhanced performance. The destabilized luc2P luciferase reporter is used for improved responsiveness to transcriptional dynamics. The luc2P gene is codon optimized for enhanced expression in mammalian cells, and the pGL4 plasmid backbone was engineered to reduce background reporter expression. The result is a cell line with very high induction levels when the pathway of interest is activated.

      Representation of the one-hybrid system.

      Example Data

      In these experiments, 10,000 transfected GloResponse™ 9XGAL4UAS-luc2P HEK293 cells per well were dispensed into each well of a 384-well plate, and serial dilutions of E2 or dexamethasone were added to induce reporter gene expression. After a 24-hour induction in a tissue culture incubator, luciferase activity was quantified using the Dual-Glo® Luciferase Assay System Reagent. n = 4 for each data point.
      E2 titration of GloResponse™ 9X<I>GAL4</I>UAS-<I>luc2P</I> HEK293 cells transfected with pBIND-ER-alpha Vector.
      E2 titration of GloResponse™ 9XGAL4UAS-luc2P HEK293 cells transfected with pBIND-ER-alpha Vector.
      Dexamethasone titration of GloResponse™ 9X<I>GAL4</I>UAS-<I>luc2P</I> HEK293 cells transfected with pBIND-GR Vector.
      Dexamethasone titration of GloResponse™ 9XGAL4UAS-luc2P HEK293 cells transfected with pBIND-GR Vector.
    1. Protocols

      Complete Protocol

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      GloResponse™ 9XGAL4UAS-luc2P HEK293 Cell Line Technical Bulletin

      PDF (630 KB)

      Quick Protocols

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      GloResponse™ 9XGAL4UAS-luc2P HEK293 Cell Line Quick Protocol FB100

      PDF (68 KB)

    2. Certificate of Analysis

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      Storage Conditions

      BELOW -140C

      Use Restrictions

      For Research Use Only. Not for Use in Diagnostic Procedures.