Application

The typical use for this product is in removing adherent cells from a culture surface. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture.

Biochem/physiol Actions

Trypsin cleaves peptides on the C-terminal side of lysine and arginine residues. The rate of hydrolysis of this reaction is slowed if an acidic residue is on either side of the cleavage site and hydrolysis is stopped if a proline residue is on the carboxyl side of the cleavage site. The optimal pH for trypsin activity is 7-9. Trypsin can also act to cleave ester and amide linkages of synthetic derivatives of amino acids. EDTA is added to trypsin solutions as a chelating agent that neutralizes calcium and magnesium ions that obscure the peptide bonds on which trypsin acts. Removing these ions increases the enzymatic activity.

Components

Trypsin consists of a single chain polypeptide of 223 amino acid residues, produced by the removal of the N-terminal hexapeptide from trypsinogen which is cleaved at the Lys - lle peptide bond. The sequence of amino acids is cross-linked by 6 disulfide bridges. This is the native form of trypsin, beta-trypsin. BETA-trypsin can be autolyzed, cleaving at the Lys - Ser residue, to produce alpha-trypsin. Trypsin is a member of the serine protease family.

Features and Benefits

StableCell^™ trypsin is formulated as a gentle solution for cell detachment. Enzyme activity is retained when stored at 2-8°C, and our studies even show >90% activity is retained when left at room temperatures for up to 7 weeks! Save Time - no more freeze or thaw cycles Save Space - store at 2-8°C and free up your freezer Does not need to be aliquoted Only the best for your cells - manufactured in GMP environment

Legal Information

StableCell is a trademark of Sigma-Aldrich Co. LLC

Preparation Note

Incubating cells with too high a trypsin concentration for a long period can damage cell membranes and kill the cells. Solubilizing trypsin or diluting it from a concentrated solution should be done with a buffered salt solution containing no Ca²⁺ or Mg²⁺.

Storage and Stability

Recommended storage is 2-8°C upon arrival. During stability studies, data showed that this product retains ≥90% of its activity when stored at 37°C for up to 8 weeks.