SRE Luciferase Reporter-HEK293 Cell Line

reporter cell line

Product Info Summary

SKU: RC1032
Size: 1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Application: Functional Assay

Product Name

SRE Luciferase Reporter-HEK293 Cell Line

SKU/Catalog Number

RC1032

Size

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

Description

The SRE Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the serum response element (SRE). The SRE reporter cell line is designed to monitor MAPK/ERK activity and can be used for studying GPCR-linked MAPK/ERK signaling pathways as well as screening of agonists, antagonists or signaling inhibitors related with the MAPK/ERK signaling pathways. Functional activity of the cell line has been validated by serum stimulation assay (Figure 1). 

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.

Cite This Product

SRE Luciferase Reporter-HEK293 Cell Line (Boster Biological Technology, Pleasanton CA, USA, Catalog # RC1032)

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 GPCR-linked MAPK/ERK signaling pathway. Screen for activators or inhibitors of the GPCR-linked MAPK/ERK 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 SRE  HEK293 cells to fetal bovine serum (FBS). 1. Harvest SRE HEK293 cells and seed cells into a white solid-bottom 96-well microplate at 5 x 10^4 cells/well in 100 µl of growth medium containing 0.5% FBS to start serum starvation. 2. Incubate cells at 37°C in a CO2 incubator for overnight. 3. The next day, stimulate cells with 40% FBS or 40% FBS plus phorbol 12-myristate 13-acetate (PMA).  4. Incubate at 37°C in a CO2 incubator for 6-16 hours.

Validation Images & Assay Conditions

Gene/Protein Information For (Source: Uniprot.Org, NCBI)

Uniprot ID

Gene ID

Gene Name

Full Name

Weight

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

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