Jan 06, 2023
Phosphate Buffered Saline (PBS)
This protocol is a draft, published without a DOI.
- 1University of Oslo
Protocol Citation: Andreas Sagen 2023. Phosphate Buffered Saline (PBS). protocols.io https://dx.doi.org/
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, phosphate buffered saline, buffered saline, phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, containing disodium hydrogen phosphate, buffer solution, potassium chloride, based salt solution, sodium chloride, salt solution, buffer, ph, pb, sodium
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
Monopotassium phosphateMerck MilliporeSigma (Sigma-Aldrich)Catalog #P5379
Sodium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9625
Potassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Disodium phosphateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9763
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++
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 8000 mg Sodium chloride, 200 mg Potassium chloride, 144 mg Disodium phosphate and 245 mg Monopotassium phosphate
Powdered compounds:
Sodium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9625
Potassium chlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #P3911
Disodium phosphateMerck MilliporeSigma (Sigma-Aldrich)Catalog #S9763
Monopotassium phosphateMerck MilliporeSigma (Sigma-Aldrich)Catalog #P5379
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 00:05:00
5m
Adjust pH while mixing to 7.4 using concentrated sodium hydroxide
Add distilled water to a total of 100 mL
Autoclave liquid at 121 °C for 00:15:00
15m