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CytoSpring

PBS (Phosphate-Buffered Saline)

320 Logue Ave, Ste 140


Mountain View, CA 94043
Email: info@cytospring.com
Tel/Fax 650-988-6699
cytospring.com

Product Information

Background Phosphate buffered saline (PBS) is a balanced salt solution commonly


used in the bio-laboratory. The essential function of a balanced salt solution is to
maintain pH and osmotic balance as well as provide cells with water and essential
inorganic ions. PBS is generally utilized to maintain cells for the short term in a viable
condition while the cells are manipulated outside of their regular growth environment.
One of the early formulas of PBS was developed by Renato Dulbecco, published in
1
1954 which are termed DPBS for Dulbeccos phosphate buffered saline.
To maintain normal function, the pH of different cellular compartments, body fluids, and
organs must be tightly regulated by a process called acid-base homeostasis. The pH of
most commonly studied biological materials fall between 7 ~ 7.6. For example, the pH of
typical cytosol is 7.2, and mitochondrial matrix is 7.5. When investigating a biological
sample under an in vitro setting, one would want to keep the condition of the sample as
close as to where its derived from. The pH value of PBS is set to be within the range of
7 ~ 7.6, and often set to be 7.4 as the pH of blood nears 7.4, a value referred to as the
physiological pH in biology and medicine. In addition to maintaining a constant pH, PBS
in general has an osmolarity that matches those of the human body (isotonic) and is
non-toxic to the cells. As a result, PBS is frequently used as a washing buffer in tissue
culture and protein chemistry, or, when combined with BSA, frequently as a diluent of
biological substances.
Storage Store the solution at room temperature or at 2 to 8C if needed.
Product The table below lists the components of DPBS and its derivatives that
CytoSpring carries. All products are sterile-filtered, suitable for cell culture and/or other
applications.

Product #
Description
[ Salt ]
NaCl
KCl
Na2HPO4
KH2PO4
CaCl2
MgCl2

P0213
DPBS, 1x
mM
137.93
2.67
8.06
1.47
-

P0243
DPBS, 10x
mM
1379.3
26.7
80.6
14.7
-

Reference
1. Dulbecco R., Vogt M. Plaque formation and isolation of pure lines with
poliomyelitis viruses. J Exp Med. 1954, 99:167182.

Q&A
Should I use PBS with or without Ca2+/Mg2+ when I wash cells?
A. It depends on your subsequent experimental procedure. Cultured cells attach the
vessel surface through the adhesion proteins which require divalent cations to function.
Many media contain calcium and magnesium which inhibit trypsin activity. Fetal bovine
serum also contains proteins that are trypsin inhibitors. One would want to use DPBS
without Ca2+/Mg2+ to wash adherent cell cultures before detaching them from the
growing surface with trypsin. Use DPBS with Ca2+/Mg2+ to wash adherent cell cultures
when you merely wish to wash and have the cells remain adherent. For Western blotting
application, there is no need of Ca2+/Mg2+ in PBS.
In addition to Ca2+/Mg2+, there are many different formulations to prepare PBS. Which
one should I use?
A. Many variations of PBS have been developed and used in the laboratory for various
applications. Originally, PBS contains the following constituents: sodium chloride,
sodium phosphate, and potassium phosphate. PBS is a physiologic buffer and normally
contains only a minor amount of potassium chloride in order to maintain membrane
potential; for special cell physiological experiments, potassium chloride may be left out.
Calcium and magnesium are included to help cell attach to surface, glucose and sodium
pyruvate, as energy and nutrient source are added on top of Ca2+/Mg2+ to maintain cells;
and thus, is sometimes called holding medium.
Follow the guideline below to choose appropriate PBS:
- Use DPBS without calcium/magnesium to wash cells if cells are to be trypsinized
subsequently.
- Use DPBS containing calcium/magnesium, glucose and sodium pyruvate if cells
are to be kept outside of incubator for a long period of time.