MTT assay is an established viability assay [35 ] and was conducted as previously described [36 (link)]. In brief, after respective treatment, MTT reagent (0.5 mg/ml MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid; Sigma-Aldrich, solved in DMEM without phenol red (GE Healthcare, München, Germany)) was applied for 2 hours at 37 °C. DMSO was added to lyse the cells. When measuring SHSY-5Y cells, this protocol was varied [37 ]. Here, MTT supernatants were collected in Eppendorf reaction tubes and centrifuged. Supernatant was discarded and pellets harvested. Pellets were re-administered to their corresponding wells, in which the cells were then lysed with DMSO. After the application of DMSO, in both approaches, the absorbance at λ = 570 nm was measured with a microplate reader (ELx800, BioTek Instruments, Bad Friedrichshall, Germany). Each MTT experiment was independently repeated 3 times.
Dmem without phenol red
Manufactured by GE Healthcare
Sourced in Germany
DMEM without phenol red is a cell culture medium that does not contain the pH indicator phenol red. It is designed for use in cell culture applications where the presence of phenol red may interfere with experimental procedures or analyses.
Automatically generated - may contain errors
2 protocols using dmem without phenol red
1
MTT Assay for Cell Viability
2
Quantifying Cell Viability with MTT Assay
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Methyl thiazolyl tetrazolium (MTT) assay [18] was performed as described previously [19] . In brief, cells were stimulated with the respective substance for the indicated time periods as described above, medium was discarded, and cells were washed with PBS, incubated for 2 h with MTT (Sigma-Aldrich/Merck, Darmstadt, Germany), and dissolved in DMEM without phenol red (GE Healthcare) (0.5 mg/mL). After removal of MTT, dimethyl sulfoxide (Roth, Karlsruhe, Germany) was added, the plates were shaken for 5 min, and adsorption was measured at 555 nm with Elx800 (BioTek, Bad Friedrichshall, Germany). Control was set as 100%.
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