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Product Info Summary
|Size:||1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.|
MRE Luciferase Reporter HEK293 Cell Line
1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
The MRE Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the metal response element (MRE). MRE is targeted by MRE-binding transcription factor-1 (MTF-1) which is a zinc finger transcription factor and plays a major role in induction of metallothionein gene expression in response to cellular stress caused by heavy metals such as zinc and cadmium. The MRE induction by ZnSO4 is shown in Figure 1.
Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Storage & Handling
Immediately upon receipt, store in liquid nitrogen.
Cite This Product
MRE Luciferase Reporter HEK293 Cell Line (Boster Biological Technology, Pleasanton CA, USA, Catalog # RC1037)
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. Actual working concentration varies and should be decided by the user.
Application:Monitor MTF-1 transcriptional activity. Screen for activators or inhibitors of the MTF-1 signaling pathway.
Culture conditions:Cells should be grown at 37°C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 3 µg/ml of Puromycin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37°C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37°C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence. To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly.
Functional validation:A. Response of MRE HEK293 cells to zinc sulfate (ZnSO4) 1. Harvest MRE HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 µl of growth medium at 5 x 104 cells/well. 2. Incubate cells at 37°C in a CO2 incubator for overnight. 3. The next day, stimulate cells with different concentrations of ZnSO4. 4. Incubate at 37°C in a CO2 incubator for 6-16 hours. 5. Add 50 µl of luciferase assay reagent per well. 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
Validation Images & Assay Conditions
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Fig-1: Induction of MRE activity by zinc sulfate in MRE HEK293 cells.
Gene/Protein Information For MRE (Source: Uniprot.Org, NCBI)
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