Activity
(Fig-1: Induction of NF-kB activity by PMA in NF-kB Leeporter™ – Jurkat T cells.)
NF-kB cell line
NF-kB Leeporter Luciferase Reporter-Jurkat Cell Line
Applications
Functional Assay
Synonyms
NF-kB; NF-kB Leeporter Luciferase Reporter-Jurkat Cell Line; NF-kB cell line
Host
Jurkat cells
Form/Format
Each vial contains 2-3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Applicable Applications for NF-kB cell line
Functional Assay (FA)
Application Notes
Monitor the NF-kB signaling pathway activity. Screen for activators or inhibitors of the NF-kB 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 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. 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 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 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.
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 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 NF-kB activity by PMA in NF-kB Leeporter™ – Jurkat T cells.)
Activity
(Fig-1: Induction of NF-kB activity by PMA in NF-kB Leeporter™ – Jurkat T cells.)
Related Product Information for NF-kB cell line
The NF-kB Leeporter Luciferase Reporter cell line is a stably transfected Jurkat T cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the NF-kB response element. NF-kB is a key transcription factor that is involved in immune and inflammatory responses, developmental processes, cellular growth and apoptosis. The NF-kB induction by phorbol 12-myristate 13-acetate (PMA) is shown in Figure 1.
Product Categories/Family for NF-kB cell line
Similar Products
Product Notes
The NF-kB (Catalog #AAA669542) is a Cell Line produced from Jurkat cells and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's NF-kB can be used in a range of immunoassay formats including, but not limited to, Functional Assay (FA). Monitor the NF-kB signaling pathway activity. Screen for activators or inhibitors of the NF-kB signaling pathway. Researchers should empirically determine the suitability of the NF-kB 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. It is sometimes possible for the material contained within the vial of "NF-kB, Cell Line" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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