TCF/LEF Luciferase Reporter-HEK293 Cell Line

Product Name

TCF/LEF Luciferase Reporter-HEK293 Cell Line

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SKU/Catalog Number

RC1019

Size

1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

Description

The TCF/LEF Luciferase Reporter cell line is a stably transfected HEK293 cell line which expresses Renilla luciferase reporter gene under the control of the TCF/LEF response element. This cell line is designed to monitor the transcriptional activity of TCF/LEF and can be used for studying Wnt signaling pathways as well as screening of activators or inhibitors that affect the TCF/LEF transcriptional activity. 

Contents

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. (Ship on dry ice.)

Cite This Product

TCF/LEF Luciferase Reporter-HEK293 Cell Line (Boster Biological Technology, Pleasanton CA, USA, Catalog # RC1019)

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 the TCF/LEF induction activity.Screen for activators or inhibitors of the TCF/LEF 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 TCF/LEF HEK293 cells to lithium chloride (LiCl) (Figure 1).1. Harvest TCF/LEF HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100  µl of growth medium at 5 x 10^4 cells/well.  2. Incubate cells at  37°C in a CO2 incubator for overnight. 3. The next day, stimulate cells with various concentrations of LiCl.  4. Incubate at  37°C in a CO2 incubator for 6-16 hours. 

Validation Images & Assay Conditions

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