ISRE cell line

ISRE Leeporter Luciferase Reporter-HEK293 Cell Line

Cat. Num. AAA669543

• Applications: Functional Assay
• Synonyms: ISRE; ISRE Leeporter Luciferase Reporter-HEK293 Cell Line; ISRE cell line

• Host: HEK293 cells
• Form/Format: Each vial contains 2-3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

• Applicable Applications for ISRE cell line: Functional Assay (FA)
• Application Notes: Monitor the JAK/STAT signaling pathway activity. Screen for activators or inhibitors of the JAK/STAT signaling pathway.
• Culture Conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium (w/ L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS and 1% Pen/Strep, plus 3ug/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 degree C water-bath, transfer to a tube containing 10ml 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 degree C-CO2 incubator. Leave the T25 flask in the incubator for 1~3 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 ISRE Leeporter - HEK293 cells to IFN-alpha.; 1. Harvest ISRE Leeporter - HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well.; 2. Incubate cells at 37 degree C in a CO2 incubator for overnight.; 3. The next day, stimulate cells with various concentrations of IFN-alpha.; 4. Incubate at 37 degree C in a CO2 incubator for 6 hours.; 5. Add 50 ul of luciferase assay reagent (Cat #MBS668917) per well.; 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
• Shipping Note: Product available only in the USA.
• Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
• Preparation and Storage: Immediately upon receipt, store in liquid nitrogen.

Activity

(Fig-1: Induction of ISRE activity by IFN-alpha in ISRE Leeporter™ – HEK293 cells.)

Related Product Information for ISRE cell line

The ISRE Leeporter Luciferase Reporter cell line is a stably transfected HEK293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the Interferon-Stimulated Response Element (ISRE), so that the cell line is designed to monitor the JAK/STAT signaling pathway activity. Functional activity of the cell line has been validated by IFN-alpha (Figure 1).

Product Categories/Family for ISRE cell line

Reporter Cell Lines