HEPES belongs to the “goods buffers” and its properties make it optimally suited as an additional buffer system in cell culture. Depending on the concentration, it stabilizes the pH value in the cell culture media in a range between 6 and 8.5. Therefore, the use of the ROTI®Cell HEPES solution is particularly recommended when the pH value urgently needs to be kept constantly high. pH fluctuations mainly occur when working outside the CO2 incubator for longer periods.
In addition to the various nutrients in the medium, the pH value also plays a very important role in cell culture. The physiological pH for almost all mammalian cells is 7.4. Deviating from this, it has been observed that certain transformed cell lines prefer a pH between 7.0 and 7.4 and certain fibroblasts prefer a pH between 7.4 and 7.7. In any case, pH fluctuations should be avoided and the pH should be maintained as well as possible. That is why a buffer system in media is necessary. The most important buffer system in the body is the carbonic acid-carbonate buffer system. Accordingly, almost all cell culture media contain bicarbonate (NaHCO3). Bicarbonate alone leads to a strong pH increase of the medium, so that NaHCO3-buffered media must always be used with CO2 incubators. Depending on the NaHCO3 concentration in the medium, different CO2 environments are necessary to ensure a pH of about 7.4. For this reason, you should not work outside of the CO2 incubator for too long. If this is necessary, a HEPES buffer should be used in addition to the NaHCO3, which contributes as a second buffer system to stabilize the pH value.
HEPES | 1 M |
pH value | 7,0-7,6 |
Osmolality | ≥1000 mOsm/kg |
Endotoxins | ≤10 EU/ml |
Sterility | complies |