TGFβ SMAD reporter assay was performed as per the manufacturer's instructions (BPS Biosciences, catalog no. 60653). Briefly, HEK293 cells were plated at a density of approximately 35,000 cells per well on to 96-well white clear-bottom plates in growth media containing MEM/EBSS (HyClone Laboratories Inc., GE Healthcare Lifesciences) with 1% non-essential amino acids (HyClone Laboratories Inc., GE Healthcare Lifesciences), 1 mmol/L sodium pyruvate (HyClone Laboratories Inc., GE Healthcare Lifesciences), 1% pen-strep (GIBCO, Thermo Fisher Scientific), and 10% FBS (MP Bio Science Ltd) and allowed to attach overnight. Spent media was discarded and cells were then incubated with recombinant TGFβ (BPS Biosciences) at 20 ng/mL and with different dilutions of BCA101, human IgG isotype antibody (R&D Systems Inc.) or TGFβRII-Fc at equimolar concentration prepared in assay media (growth media containing 0.5% FBS). After 18 hours, cells were incubated with 100 μL of BioGlo luciferase reagent for 15 minutes at room temperature. Luminescence readout was taken as per manufacturer's instructions using a Spectramax M5e plate reader (Molecular Devices) with SoftMax Pro GxP software version 6.5. EC50 was analyzed using SoftMax Pro software and data plotted using GraphPad Prism software version 9.
Mem ebss
Manufactured by Cytiva
Sourced in United States
MEM/EBSS is a powdered cell culture medium that provides essential nutrients and salts for the growth and maintenance of various cell types in vitro. It is a balanced salt solution that supports cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Penicillin streptomycin neomycin, by Thermo Fisher Scientific (2 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by GE Healthcare (1 mentions)
Fetal bovine serum (fbs), by R&D Systems (1 mentions)
Non essential amino acids (neaa), by GE Healthcare (1 mentions)
Spectramax m5e plate reader, by Molecular Devices (1 mentions)
3 protocols using mem ebss
1
TGFβ SMAD Reporter Assay in HEK293 Cells
2
IMR-90 Fibroblast Culture on SESM Acrylamide Gel
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cell culture IMR-90 lung fibroblast cells were obtained from ATCC and maintained in minimum essential media with Earle's balanced salts (MEM/EBSS; Hyclone Laboratories Inc, Logan, UT, USA) supplemented with 10% fetal bovine serum (Sigma, St. Louis, MO, USA), sodium pyruvate (Gibco, Waltham, MA, USA), non-essential amino acids (NEAA; Gibco), and penicillin-streptomycin-neomycin (PSN; Gibco). IMR-90 (passage 3, 1 x 10 5 cells) were seeded in a 10 cm diameter control tissue culture dish (n = 3) and a tissue culture dish containing a thin acrylamide gel (8 cm diameter) with SESM bound to it (n=3) using a method described previously. The cells were cultured for 51 h.
3
IMR-90 Fibroblast Culture on SESM Acrylamide Gel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell culture IMR-90 lung fibroblast cells were obtained from ATCC and maintained in minimum essential media with Earle's balanced salts (MEM/EBSS; Hyclone Laboratories Inc, Logan, UT, USA) supplemented with 10% fetal bovine serum (Sigma, St. Louis, MO, USA), sodium pyruvate (Gibco, Waltham, MA, USA), non-essential amino acids (NEAA; Gibco), and penicillin-streptomycin-neomycin (PSN; Gibco). IMR-90 (passage 3, 1 x 10 5 cells) were seeded in a 10 cm diameter control tissue culture dish (n = 3) and a tissue culture dish containing a thin acrylamide gel (8 cm diameter) with SESM bound to it (n=3) using a method described previously. The cells were cultured for 51 h.
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