Trypsin Solution (1X)
Contains 0.25% porcine trypsin (1:250) in HBSS without Ca2+ and Mg2+
With phenol red
Sterile-filtered
Porcine parvovirus tested
Cell culture tested
Endotoxin tested

Catalog Number LS 015-03
Store at less than 0°C

Product Description
In adherent cell culture, Trypsin or Trypsin-EDTA (ethlyenediaminetetraacetic acid) is used for the detachment of cell adhesion from culture dish or flask, and degradation of cell-cell adhesion. Trypsin, a proteolytic enzyme, can cleave collagen that is pre-coated on culture surface for cell adhesion. EDTA, a chelating agent, can bind with cations such as Ca2+ and Mg2+ which are involved in cell adhesion. Subculture usually requires chelation of cations and proteolytic reaction, but the severity of the treatment (high concentration or long time) will reduce the viability and the rate of cell growth because of the over-degradation of cell surface proteins and deficiency of cations essential for cell. A protocol should be selected with the least severity compatible with the generation of a single cell suspension of high viability.

LS 015-03 (1X) contains 0.25% (w/v) procine trypsin (LS 015-05) in Hanks’ balanced salt solution (HBSS, LB 003-03). The concentration of Trypsin is strongly influenced by (1) type of cell, (2) type of culture, and (3) characteristics of cell growth. It is important to review the literature for recommendations concerning medium, supplementation and physiological parameters required for a specific cell line.

Storage/Stability
Trypsin solutions should be stored at -5~-20°C in the dark, and can be stored at 2~8°C for 1~2 weeks. Deterioration of the solution may be recognized by (1) precipitate or particulate matter throughout the solution, (2) cloudy appearance, (3) color change, (4) pH change and/or (5) temperature change. Product label bears expiration date.

Biological Performance Characteristics
The detaching and growth-maintaining capacities of Trypsin are tested in a medium supplemented with 10% FBS using an appropriate adherent cell line(s). Growth rates are examined through three subculture generations and compared with parallel cultures grown in standardized control medium. Cells are counted and growth is plotted as a logarithmic function of time in culture, and seeding efficiency, doubling time, and final cell densities are determined. During the testing period cultures are examined microscopically for a typical morphology and evidence of cytotoxicity.

For Use

1. Remove all medium from culture vessel.
2. Wash the cell monolayer with Ca2+ and Mg2+ -free salt solution, D-PBS (LB 001-02) or HBSS (LB 003-03). Remove salt solution.
3. Dispense proper volume of Trypsin solution to culture vessel to cover the cell monolayer of cells completely.
4. Remove the Trypsin solution and replace the closed culture vessel in incubator at 37°C for 2-5 minutes.
5. Check a progress with an inverted microscope.
6. Cells can be resuspended by gently pipetting with medium containing serum. Serum inhibits further tryptic activity.

Precautions
For In Vitro Use Only