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20 mM HEPES pH 7.4

Working buffer for live-cell imaging and electrophysiology.

Component100 mL500 mL1 L
HEPES (free acid)0.4766 g2.383 g4.766 g
NaOH (1 M)approximate; finish with a pH meter0.9082 mL4.541 mL9.082 mL
Final volume with water100 mL500 mL1 L
  1. Dissolve HEPES (free acid) in about 80 % of the final volume of purified water.
  2. Adjust to pH 7.4 at 25 °C with NaOH (1 M), adding slowly while stirring.
  3. Make up to the final volume with water.
  4. Filter-sterilise (0.22 µm) and store at 4 °C.