NFAT Reporter – Hek293 Cell Line (PKC/ Ca2+ Pathway)

Catalog #: 79298
Size: 2 Vials
The protein kinase c (PKC)/ Ca2+ response pathway leads to activation of the transcription factor nuclear factor of activator T cells (NFAT). NFAT is regulated by Ca2+ and the Ca2+/calmodulin-dependent serine phosphatase calcineurin. NFAT proteins are phosphorylated and reside in the cytoplasm in resting cells; upon stimulation, they are dephosphorylated by calcineurin, translocate to the nucleus, and induce gene expression.
The NFAT Reporter – Hek293 cell line contains a firefly luciferase gene under the control of NFAT response element stably integrated into Hek293 cells. This cell line is validated for the response to the stimulation of phorbol 12-myristate 13-acetate (PMA) with ionomycin.
Materials Required But Not Provided:

• PMA (LC Laboratories, #P1680) Prepare stock solution in DMSO.
• Ionomycin (Sigma, #I3909) Prepare stock solution in DMSO. 
• Assay medium: Thaw Medium 1 (BPS Bioscience, #60187)
Growth Medium 1B (BPS Cat. #79531)
• 96-well tissue culture plate or 96-well tissue culture-treated white clear-bottom assay plate 
• RO-31-8220 (EMD Millipore, #557520) or other PKC inhibitor
ONE-Step Luciferase Detection Reagents (BPS Bioscience, #60690) for measuring firefly luciferase activity.
• Luminometer 

Nuclear factor of activated T cells; T cell, PKC Ca 2+, Calcium PKC Pathway
Supplied As:
Each vial contains ~2 X 10^6 cells in 1 ml of 10% DMSO.
Host Cell line:
Mycoplasma Testing:
The cell line has been screened using the PCR-based VenorGeM® Mycoplasma Detection kit (Sigma-Aldrich, #MP0025) to confirm the absence of Mycoplasma species.
Growth Media:

Thaw Medium 1 (BPS Cat. #60187): MEM medium (Hyclone #SH30024.01) supplemented with 10% FBS (Invitrogen #26140-079), 1% non-essential amino acids (Hyclone #SH30238.01), 1 mM Na pyruvate (Hyclone #SH30239.01), 1% Penicillin/Streptomycin (Hyclone SV30010.01).

Growth Medium 1B (BPS Cat. #79531): Thaw Medium 1 plus 400 μg/ml of Geneticin (Life Technologies #11811031).

Instructions for use:
See data sheet for detailed culturing and assay protocol.
Storage / Stability:
Immediately upon receipt, store in liquid nitrogen.
Monitor intracellular calcium levels, and screen for activators or inhibitors of the PKC/ Ca2+ pathway.

License Disclosure: Purchase of this cell line grants you with a 10-year license to use this cell line in your immediate laboratory, for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make the cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit the use of this cell line in humans or for therapeutic or drug use. The license does not permit modification of the cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license and result in an immediate cease of sales and distribution of BPS products to your laboratory. BPS does not warrant the suitability of the cell line for any particular use, and does not accept any liability in connection with the handling or use of the cell line. Modifications of this cell line, transfer to another facility, or commercial use of the cells may require a separate license and additional fees; contact for details. Publications using this cell line should reference BPS Bioscience, Inc., San Diego.

Avoid freeze/thaw cycles.
Scientific Category:
Cell Signaling Pathway
Product Type:
Cell Line
Data shown is lot-specific. Contact us for specific information on other lots.

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