STAT4 cell line

STAT4 Leeporter Luciferase Reporter-Ba/F3 Cell Line

Cat. Num. AAA669545

• Applications: Functional Assay
• Synonyms: STAT4; STAT4 Leeporter Luciferase Reporter-Ba/F3 Cell Line; STAT4 cell line

• Host: Ba/F3 cells
• Form/Format: Each vial contains 2-3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

• Applicable Applications for STAT4 cell line: Functional Assay (FA)
• Application Notes: Monitor the STAT4 signaling pathway activity. Screen for activators or inhibitors of the STAT4 signaling pathway.
• Culture Conditions: Cells should be grown at 37 degree C with 5% CO2 using RPMI medium supplemented with 10% heat-inactivated FBS, 1 mM sodium pyruvate, 10 mM HEPES and 1% Pen/Strep, plus 5 ng/ml mIL-3 (Note: mIL-3 is essential for Ba/F3 cell maintenance), plus 3ug/ml of Puromycin. (Note: The parental Ba/F3-Puro cell line (Part #14135CCL) is a blank vector-transfected stable cell line which also requires 3ug/ml of Puromycin for cell culture maintenance! 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. Monitor the cell viability by counting cells daily for 1~3 days until cells completely recover viability as cells are doubling daily. Once cells are over 90% confluent, harvest cells by centrifugation and passage cells. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence. To passage the cells, transfer cells to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cell 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 STAT4 Leeporter - Ba/F3 cells to mIFN-gamma.; 1. Harvest STAT4 Leeporter - Ba/F3 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium without IL-3 at 1 x 10^5 cells/well.; 2. Right after plating cells, stimulate cells with various concentrations of mouse IFN-gamma and incubate cells at 37 degree C in a CO2 incubator for 6-16 hours.; 3. Add 50 ul of luciferase assay reagent (Cat #MBS668917) per well.; 4. 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 STAT4 activity by mIFN-gamma in STAT4 Leeporter™ – Ba/F3 cells.)

Related Product Information for STAT4 cell line

The STAT4 Leeporter Luciferase Reporter cell line is a stably transfected Ba/F3 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the STAT4 responsive promoter, so that the cell line is designed to measure the transcriptional activity of STAT4. Signal Transducer and Activator of Transcription 4 (STAT4) is a member of the STAT transcription factor family and plays a central role in generating inflammation during protective immune responses and immune-mediated diseases. The STAT4 induction by interferon gamma is shown in Figure 1.

Product Categories/Family for STAT4 cell line

Reporter Cell Lines