Preparation HEPES Buffer Saline (HBS)

This guide describes the preparation of HEPES Buffer Saline (HBS) at a defined molarity for laboratory use.

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HEPES Buffer Saline (HBS) is commonly used in transfection experiments and cell culture applications.

HEPES Buffer Saline (HBS) – Composition

NameFormulaConcentrationCAS
Sodium chlorideNaCl135 mM7647-14-5
Potassium chlorideKCl5 mM7447-40-7
Disodium phosphateNa₂HPO₄750 µM7778-77-0
GlucoseC₆H₁₂O₆5 mM50-99-7
HEPESC₈H₁₈N₂O₄S20 mM7365-45-9

Sterilization & Storage

  • Sterilization: 0.22 μm filtration.
  • Storage: Aliquot and store at −20 °C.

Tips

  • Before use in transfection experiments, test the preparation by mixing 0.5 mL of the buffer with 0.5 mL of 250 mM CaCl₂ and vortexing. A fine precipitate should form and be visible under a microscope. If no precipitate forms, do not use this batch for transfection.
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