The SBE Reporter – HEK293 Cell Line is designed for monitoring the activity of the TGF /SMAD signaling pathway. The transforming growth factor beta (TGF) signaling pathway is involved in a diverse range of cell processes such as differentiation, cell cycle arrest, and immune regulation. TGF signaling has been linked to cardiac disease, cancer, Alzheimer’s and other human diseases. TGF proteins bind to receptors on the cell surface, initiating a signaling cascade that leads to phosphorylation and activation of SMAD2 and SMAD3, which then form a complex with SMAD4. The SMAD complex then translocates to the nucleus and binds to the SMAD binding element (SBE) in the nucleus, leading to transcription and expression of TGF / SMAD responsive genes.

Human TGFβ1 (BPS Cat. #90900-1)

SB525334 (Selleckchem #356559-20-1): inhibitor of TGFβ pathway. Prepare stock solution in DMSO.

Thaw Medium 1 (BPS Bioscience #60187)

Growth Medium 1B (BPS Bioscience #79531)

Assay Medium 1B (BPS Bioscience #79617)

96-well tissue culture-treated white clear-bottom assay plate (Corning # 3610)

ONE-Step™ Luciferase Assay System (BPS Cat. #60690)

Luminometer

SBE cell line, TGFbeta reporter, SMAD reporter, luciferase reporter

Each vial contains ~1.5 X 10⁶ cells in 1 ml of 10% DMSO.

This cell line is validated for its response to the stimulation of human TGFβ1 and to treatment with an inhibitor of the TGFβ/SMAD signaling pathway.

HEK293 cells

The cell line has been screened using the PCR-based VenorGeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.

Thaw Medium 1 (BPS Cat. #60187): MEM medium (Hyclone #SH30024.01) + 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 (BPS Cat. #60187) plus 400 µg/ml of Geneticin (Invitrogen #11811031).

Cells should be grown at 37°C with 5% CO2 using Growth Medium 1B.

Aqueous solution containing DMSO.

See data sheet for detailed culturing and assay protocol.

Store in liquid nitrogen immediately upon receipt.

• Monitor TGF signaling pathway activity.
• Screen for activators or inhibitors of the TGF/SMAD signaling pathway.

Moustakas A et al. (2001) Smad regulation in TGF-beta signal transduction. J.Cell Science. 114(Pt 24): 4359-69.

Application Reference(s):
1. Functional Adenosine Receptor 2B Drives Profibrotic and Proinflammatory Signaling in Renal Fibroblasts Through Modulation of TGFβ Receptor Signaling (2017)

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 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 sales@bpsbioscience.com for details. Publications using this cell line should reference BPS Bioscience, Inc., San Diego.

Avoid freeze/thaw cycles

Cell Signaling Pathway

Cell Line