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EBSS (magnesium, calcium, phenol red) (pH 7.0) Preparation and Recipe

EBSS is the abbreviation of Earle's Balanced Salt Solution, and is in common use for cell culture applications including washing and dilution in CO2 environments. The addition of the magnesium and calcium ions, along with phenol red, is to assist in reagent preparation. Phenol red is a weak estrogen and should not be used long-term with responsive cell cultures. For other buffers containing Phenol Red, click here.

To prepare  L of EBSS (magnesium, calcium, phenol red) (pH 7.0):
Change the value in the textbox above to scale the recipe volume

Table 1. Required components
Calcium Chloride (mw: 110.98 g/mol)0.2 g0.001 M
Magesium Sulfate Heptahydrate (mw: 246.47 g/mol)0.2 g0.00081 M
Potassium Chloride (mw: 74.55 g/mol)0.4 g0.005 M
Sodium bicarbonate (mw: 84.01 g/mol)2.2 g0.026 M
Sodium chloride (mw: 58.44 g/mol)6.8 g0.117 M
Sodium Phosphate Monobasic Monohydrate (mw: 137.99 g/mol)0.14 g0.001 M
D-Glucose (Dextrose) (mw: 180.16 g/mol)1 g0.006 M
Phenol Red (mw: 354.38 g/mol)0.01 g0.00003 M
  1. Prepare 800 mL of distilled water in a suitable container.
  2. Add 0.2 g of Calcium Chloride to the solution.
  3. Add 0.2 g of Magesium Sulfate Heptahydrate to the solution.
  4. Add 0.4 g of Potassium Chloride to the solution.
  5. Add 2.2 g of Sodium bicarbonate to the solution.
  6. Add 6.8 g of Sodium chloride to the solution.
  7. Add 0.14 g of Sodium Phosphate Monobasic Monohydrate to the solution.
  8. Add 1 g of D-Glucose (Dextrose) to the solution.
  9. Add 0.01 g of Phenol Red to the solution.
  10. Adjust solution to final desired pH using HCl or NaOH
  11. Add distilled water until the volume is 1 L.

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Physiological Buffer
pH Buffering
Sample Preparation
Cell/Culture/Growth Media
Gel Electrophoresis


This online tool may be cited as follows


"Quest Calculate™ EBSS (magnesium, calcium, phenol red) (pH 7.0) Preparation and Recipe." AAT Bioquest, Inc.28 May2024https://www.aatbio.com/resources/buffer-preparations-and-recipes/ebss-calcium-magnesium-phenol-red-ph-7.


AAT Bioquest, Inc. (2024May 28). Quest Calculate™ EBSS (magnesium, calcium, phenol red) (pH 7.0) Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/ebss-calcium-magnesium-phenol-red-ph-7.