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Ciprofloxacin HydrochlorideCiprofloxacin hydrochloride (HCl) is a fluoroquinolone anti-biotic. 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 InformationMycoplasma 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 contami-nation, 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; there-fore regular feeding schedules are not interrupted. Treat-ment 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.

Working ConcentrationThe 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

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 HClCiprofloxacin 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 media.

Ciprofloxacin HCl may be used in combination with other antibiotics.

Ciprofloxacin HCl is soluble in water. Stock concentra- tions of up to 10 mg/mL can be made and stored frozen.

Cat. No. Description Size Qty/Pk

61-277-RF Ciprofloxacin HCl 1 g 1

61-277-RG Powder 5 g 1

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