NF-kB Leeporter Luciferase Reporter cell line product blog
Tags: Cell Line; NF-kB Leeporter Luciferase Reporter; NF-kB Leeporter Luciferase Reporter cell line;
The NF-kB Leeporter Luciferase Reporter n/a (Catalog #MBS669106) is a Cell Line and is intended for research purposes only. The product is available for immediate purchase. MyBioSource\'s NF-kB Leeporter Luciferase Reporter can be used in a range of immunoassay formats including, but not limited to, Functional Assay.- Monitor the NF-kB signaling pathway activity.
- Screen for activators or inhibitors of the NF-kB signaling pathway.
Functional validation:
A. Response of NF-kB Leeporter™ � Jurkat cells to phorbol 12-myristate 13-acetate (PMA).
1. Harvest NF-kB Leeporter™ � Jurkat cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 2.5 x 10^5 cells/well.
2. Right after plating cells, stimulate cells with various concentrations of PMA and incubate cells at 37�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. Researchers should empirically determine the suitability of the NF-kB Leeporter Luciferase Reporter n/a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process.
To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
Please refer to the product datasheet for known applications of a given cell line. We\'ve tested the NF-kB Leeporter Luciferase Reporter-Jurkat Cell Line with the following immunoassay(s):
Testing Data (Fig-1: Induction of NF-kB activity by PMA in NF-kB Leeporter Jurkat T cells.)
The NF-kB Leeporter� Luciferase Reporter cell line is a stably transfected Jurkat T cell line which expresses Renilla luciferasereporter gene under the transcriptional control of the NF-kB response element. NF-kB is a key transcription factor that isinvolved in immune and inflammatory responses, developmental processes, cellular growth and apoptosis. The NF-kB inductionby phorbol 12-myristate 13-acetate (PMA) is shown in Figure 1.
Culture conditions: Cells should be grown at 37�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 3 ?g/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.
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. Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.