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Ciprofloxacin Hydrochloride

Ciprofloxacin hydrochloride (HCl) is a fluoroquinolone antibiotic. It is effective against a wide range of gram positive and
gram negative bacteria1, and is most well known for its
effectiveness against mycoplasma. Ciprofloxacin HCl works
by interfering with the bacterial enzyme DNA gyrase, an
enzyme necessary for bacterial synthesis, replication, and
transcription in both the active and non-active growth phases
of the bacterial life cycle.

Background Information
Mycoplasma contamination may originate from various
sources including cell culture medium, serum added to the
medium, tissue used to establish primary cultures, and most
commonly is passed from individuals who handle the cultures.
Mycoplasma is an insidious infection that decreases cellular
activity and growth rate4, produces membrane alterations5,
affects amino acid6 and nucleic acid metabolism, causes
chromosome aberrations7, and diminishes the quality of
resultant data or products. Mycoplasma infections often
remain undetectable and even with the heaviest contamination, cultures may appear to grow normally and remain
clear. Reports estimate that up to 35% of cultures containing
mycoplasma go undetected8. The most common method
for detecting mycoplasma is the Barile and Kern method. If
detected, Ciprofloxacin HCl can provide the end user with
an alternative to discarding valuable cell cultures, thus saving
valuable time and money. Ciprofloxacin HCl shows minimal
decrease in effectiveness even after six days in culture; therefore regular feeding schedules are not interrupted. Treatment can be discontinued when no signs of reoccurrence are
apparent after four consecutive weeks9. Ciprofloxacin HCl
and the protocols for its use are patented- US patent numbers
4,670,444 and 4,895,803.

well as several species of mycoplasma. In comparison to other

antibiotics, Ciprofloxacin HCl has many advantages, including
lack of known resistant strains, minimal side effects, a lower
effective working concentration, non-cytotoxicity, a low
reoccurrence rate for contamination, and the ability to kill
bacteria in both the active and non-active growth phases.

Working with Ciprofloxacin HCl

Ciprofloxacin HCl powder should be stored at room
temperature below 30C. Liquid stock solutions should
be stored frozen (-20C).
To prepare a stock solution add the appropriate amount
of powder to distilled deionized water while mixing.
NOTE: Adding water directly to the powder may result in
insoluble mixtures that cannot be filtered without
losing potency. After ensuring that the mixture is
completely solubilized, filter sterilized. Store at -20C.
If desired, the pH may be adjusted.
Media containing freshly added Ciprofloxacin HCl, at
an effective working concentration of 5-25 g/mL (most
commonly 10 g/mL), should be added to cells seeded at a
density of 10 to 105.
Cells should be re-seeded or sub-cultured with media
containing Ciprofloxacin HCl at 3 to 4 day intervals.
Cells should be treated for 12-20 days, after which, it is
no longer necessary to include Ciprofloxacin HCl in the
Ciprofloxacin HCl may be used in combination with other
Ciprofloxacin HCl is soluble in water. Stock concentrations of up to 10 mg/mL can be made and stored frozen.

The effective concentration for Ciprofloxacin HCl, as with

many antibiotics varies with the organism or the cell type,
environmental conditions, and stage of growth cycle.
However, a concentration of 10 g/mL is active against
most strains of gram negative and gram positive bacteria, as

Cat. No.





Ciprofloxacin HCl





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Working Concentration