NFAT Leeporter™ Luciferase Reporter-RAW264.7 Cell Line
Fig-1: Induction of NFAT activity by calcium ionophore A23187 in NFAT Leeporter™ - RAW264.7 cells.
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Order now and get it on Wednesday November 06, 2024
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| Amount : | 1 Vial |
| Content : | Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO. |
| Storage condition : | Immediately upon receipt, store in liquid nitrogen. |
The NFAT Leeporter™ Luciferase Reporter cell line is a stably transfected RAW264.7 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the Nuclear Factor of Activated T-cells (NFAT) response element, so that the cell line is designed to measure the transcriptional activity of NFAT. NFAT is a transcription factor originally found in activated T lymphocytes, and is now known to regulate not only T cell activation and differentiation but also the function of other immune cells including dendritic cells, B cells and megakaryocytes. The NFAT induction by calcium ionophore A23187 is shown in Figure 1.
Application:
- Monitor the NFAT signaling pathway activity.
- Screen for activators or inhibitors of the NFAT signaling pathway.
Culture conditions:
Cells should be grown at 37oC 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 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 37oC 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 37oC-CO2 incubator.
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 NFAT Leeporter™ – RAW264.7 cells to calcium ionophore A23187.
LIMITED USE RESTRICTIONS:
THIS PRODUCT IS SOLELY FOR IN VITRO RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE.
By use of this product, user agrees to be bound by the terms of this limited use statement.
This product is solely for Internal Research Purposes and not for Commercial Purposes. Commercial Purposes include, but are not limited to (1) use of the cell line in manufacturing; (2) use of the cell line to provide a service, information or data; (3) use of the cell line for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the cell line whether or not such cell lines are resold for use in research. The buyer cannot sell, give or otherwise transfer this product to a third party.
Commercial License Agreement is available for non-research use if applicable. Please contact Abeomics ([email protected]).
For Research Use Only. Not for use in diagnostic/therapeutics procedures.
Karpurapu M, Nie Y, Chung S, Yan J, Dougherty P, Pannu S, Wisler J, Harkless R, Parinandi N, Berdyshev E, Pei D and Christman JW (2024)
The calcineurin–NFATc pathway modulates the lipid mediators in BAL fluid extracellular vesicles, thereby regulating microvascular endothelial cell barrier function.
Front. Physiol. 15:1378565. doi: 10.3389/fphys.2024.1378565
https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2024.1378565/full
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