AP-1 Luciferase Reporter-HEK293 Cell Line

Product Name

AP-1 Luciferase Reporter-HEK293 Cell Line

SKU/Catalog Number

RC1002

Size

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

Description

The AP-1 Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the activator protein 1 (AP-1). The AP-1 transcription factors are homo- or  hetero-dimers that consist of proteins belonging to a group of structurally and functionally related members of the Jun family (c-Jun, JunB and JunD), the Fos (c-Fos, FosB, Fra-1 and Fra-2) and the subfamilies of ATF (ATFa, ATF-2 and ATF-3) and JDP (JDP-1 and JDP-2).  The AP-1 induction by phorbol 12-myristate 13-acetate (PMA) is shown in 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. (Ship on dry ice.)

Cite This Product

AP-1 Luciferase Reporter-HEK293 Cell Line (Boster Biological Technology, Pleasanton CA, USA, Catalog # RC1002)

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	Dilution	Species
Screen for activators or inhibitors of the AP-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 (Note: Puromycin can be omitted during the reporter cell assays).
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.To achieve satisfactory results, cells should not be passaged over 16 times. Functional validation: A. Response of AP-1  HEK293 cells to phorbol 12-myristate 13-acetate (PMA)
3. The next day	stimulate cells with different concentrations of PMA.

Validation Images & Assay Conditions

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