What could be the reason and will it effect the growth of my cell line? Gen5™ Microplate Reader and Imager Software, besides controlling reader function, can be used to calculate pH from previously established pH calibration curves. Le rouge de phénol (également connu sous les noms de phénolsulfonephtaléine ou PSP) est un indicateur coloré utilisé en chimie dans les dosages acidobasiques.Son pKa est de 8,0 ; sa forme acide est jaune ; sa forme basique est rouge. -Sodium Pyruvate is present in DMEM with phenol red and absent in DMEM without phenol red. Figure 10. If you remove the high CO2, the sodium bicarbonate takes over and you get a … Dulbecco's Modified Eagle Medium (DMEM), 500mL, Liquid, 1X Concentration, 7.0 to 7.4 pH, With High Glucose, L-Glutamine, Phenol Red, 12 Months Shelf Life, 320 to 360mOsm/kg Osmolality, For Mammalian Cell Culture These mediums generally add some non-essential amino acids and vitamins, including serine, proline, biotin, Vitamin B12, etc. The incubator does not have CO2 control. BioTek is a family-run organization with origins dating back to 1968. In solution at very low pH phenol red is colorless and exists as a zwitterion, containing both a negatively charged sulfate group and a positively charged ketone group. This acts as a zwitterion and has proven superior to conventional buffers in comparative biological assays with cell-free preparations. These increases are consistent across the spectrum and as such, ratiometric analysis or background subtraction will be corrective. Is there any specific culture media for SH-SY5Y cell lines? Thus, at 25°C, HEPES has a pK = 7.55 with ΔpK/°C = -0.014, MOPS has a pK= 7.20 with ΔpK/°C = -0.013, and BES has a pK = 7.15 with ΔpK/°C = -0.016. A smaller peak is observed at 415 nm with acidic pH levels that diminishes with an increase in pH. You need CO2 for proper buffering of DMEM. Search or browse our database to find scientific articles submitted by BioTek’s applications scientists and published in leading journals. Specific supplement concentrations are assay dependent. If you are not currently subscribed, you may sign up now to begin receiving issues as they are released. The absorbance of complete DMEM media was measured at 415 nm and 560 nm at various pH levels and the ratio of the two values plotted as a function of pH. DMEM, which contains the most phenol red dye had the greatest change, but the relative fold-increase for the different media is the same (approx. Cat. The BioTek Global Technical Support Center (GTS) is your Help Desk for all questions related to BioTek product use, maintenance, troubleshooting support and other frequently asked questions. Most mammalian tissues exist at a near neutral pH. This year we celebrate 50 years of Passion & Innovation. Likewise the transfer of a plate to and from the reader has the potential to result in condensation of the under surface of the plate lid. DMEM was originally formulated with low glucose (1 g/L) and sodium pyruvate, but is often used with higher glucose levels, with or without sodium pyruvate. Check back often for our latest additions to the Visual Abstract library! HyClone DMEM media preparations are available in high or low glucose formulations, as well as with or without L-glutamine, magnesium, and sodium pyruvate. Do you have a suggestion about what to do? Carbon dioxide dissolves into the media forming carbonic acid as it reacts with water. DMEM was originally formulated with low glucose (1 g/L) and sodium pyruvate, but is often … Additionally, the formulation also includes glycine, serine and ferric nitrate. We do keep our cultures in 10% CO2 but for experiments they remain outside incubator for 30-40 minutes. For experiments, cells were plated into Corning 3904 black sided clear bottom 96-well microplates. DMEM (44mM NaHCO3, orange line) is held at physiological pH at CO2 concentrations between 7.5% and 11%. Wavelength with absorbance peak in the presence (+) or absence (-) of phenol red is indicated with an arrow. The formulation of glucose in the media is also altered. Paul lucas is on the spot. This documents contains … Hi, beside the discussion of Co2 I would first check if DMEM is the right medium for culturing Hepatocytes. It is true that mixing together three buffers at identical concentrations is often a good procedure to cover a broad range of operational pH's and, as stated, gives a total concentration three times that of a single one.