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

Component	Amount	Concentration
Calcium Chloride (mw: 110.98 g / mol)	#	#
Magesium Sulfate Heptahydrate (mw: 246.47 g / mol)	#	#
Potassium Chloride (mw: 74.55 g / mol)	#	#
Sodium bicarbonate (mw: 84.01 g / mol)	#	#
Sodium chloride (mw: 58.44 g / mol)	#	#
Sodium Phosphate Monobasic Monohydrate (mw: 137.99 g / mol)	#	#
D-Glucose (Dextrose) (mw: 180.16 g / mol)	#	#
Phenol Red (mw: 354.38 g / mol)	#	#

Prepare 800 mL of distilled water in a suitable container.
Add # of Calcium Chloride to the solution
Add # of Magesium Sulfate Heptahydrate to the solution
Add # of Potassium Chloride to the solution
Add # of Sodium bicarbonate to the solution
Add # of Sodium chloride to the solution
Add # of Sodium Phosphate Monobasic Monohydrate to the solution
Add # of D-Glucose (Dextrose) to the solution
Add # of Phenol Red to the solution
Adjust solution to final desired pH using HCl or NaOH
Add distilled water until the volume is 1 L.

References

This online resource may be cited as follows

MLA	"Quest Calculate™ EBSS (magnesium, calcium, phenol red) (pH 7.0) Preparation and Recipe." AAT Bioquest, Inc., 7 Oct. 2025, https://www.aatbio.com/resources/buffer-preparations-and-recipes/ebss-calcium-magnesium-phenol-red-ph-7.
APA	AAT Bioquest, Inc. (2025, October 7). 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.
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