Jan 06, 2023

Public workspacePhosphate Buffered Saline (PBS)

This protocol is a draft, published without a DOI.
  • 1University of Oslo
Andreas Sagen
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Protocol Citation: Andreas Sagen 2023. Phosphate Buffered Saline (PBS). protocols.io https://protocols.io/view/phosphate-buffered-saline-pbs-cmdqu25w
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 06, 2023
Last Modified: January 06, 2023
Protocol Integer ID: 74896
Keywords: Buffer, PBS, phosphate buffered saline
Abstract
Phosphate-buffered saline (PBS) is a buffer solution (pH ~7.4) commonly used in biological research. It is a water-based salt solution containing disodium hydrogen phosphate, sodium chloride and optionally, potassium chloride and potassium dihydrogen phosphate. The buffer helps to maintain a constant pH. PBS has many uses because it is isotonic and non-toxic to most cells[1].
Guidelines
Follow step by step, unless stated otherwise. Equipment needed should be standard to a microbiology lab.
Materials
Analytical scale, autoclave, bottle, weight vessel, LAF bench
Protocol materials
ReagentSodium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9625
Step 1.2
ReagentPotassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Step 1.2
ReagentDisodium phosphateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9763
Step 1.2
ReagentMonopotassium phosphateMerck MilliporeSigma (Sigma-Aldrich)Catalog #P5379
Step 1.2
Safety warnings
When removing autoclaved components, be sure to take care as this can be very hot.
Before start
Prepare glassware by cleaning it, and ensure that scale is sufficiently calibrated
100 mL 10x PBS w/o Ca2++, Mg2++
100 mL 10x PBS w/o Ca2++, Mg2++
20m
20m
All compounds are measured using a high precision analytical scale from powdered compounds. Each compound is measured to within 1% of the target weight. All compounds are mixed in a Duran bottle
Fill the bottle with 50 mL double-distilled water
Measure Amount8000 mg Sodium chloride, Amount200 mg Potassium chloride, Amount144 mg Disodium phosphate and Amount245 mg Monopotassium phosphate Powdered compounds: ReagentSodium chlorideSigma AldrichCatalog #S9625
ReagentPotassium chlorideSigma AldrichCatalog #P3911
ReagentDisodium phosphateSigma AldrichCatalog #S9763
ReagentMonopotassium phosphateSigma AldrichCatalog #P5379
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for Duration00:05:00
5m
Adjust pH while mixing to Ph7.4 using concentrated sodium hydroxide
Add distilled water to a total of Amount100 mL
Autoclave liquid at Temperature121 °C for Duration00:15:00

15m