High-Glucose DMEM with L-Glutamine and HEPES

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• DMEM (Dulbecco's Modified Eagle Medium)DMEM (Dulbecco's Modified Eagle Medium) is a basal cell culture medium derived from Eagle's Minimal Essential Medium by raising the amino acid and vitamin concentrations roughly fourfold. It is supplied in high-glucose (4,500 mg/L, 25 mM) and low-glucose (1,000 mg/L, 5.6 mM) forms, buffered with 3,700 mg/L sodium bicarbonate, and requires serum or a defined supplement plus a CO2 atmosphere to hold physiological pH. DMEM is the default medium for adherent lines such as HEK293, HeLa, NIH/3T3, Vero and CHO-derived adherent cultures, and for most primary fibroblasts.
• Phenol Red in Cell Culture MediaPhenol red (phenolsulfonphthalein) is a pH indicator added to cell culture media at roughly 5-15 mg/L, where it turns yellow below about pH 6.8 and pink to fuchsia above about pH 8.2, with the normal red-orange of healthy medium sitting near pH 7.4. Medium turning yellow means acidification - almost always lactate from cell metabolism, from an overgrown culture or from bacterial contamination - while medium turning pink or purple means the medium has gone alkaline, nearly always because CO2 has escaped from the bicarbonate buffer. Phenol red does no buffering itself and is purely diagnostic; use phenol-red-free medium for fluorescence and absorbance assays, and for oestrogen-responsive cell work, where the dye interferes with the readout.