胰酶 Trypsin (0.25%), EDTA (0.02%)

Product Description:

Trypsin, an animal-derived product, is the most commonly used enzyme for harvesting cells in culture. Trypsin is a pancreatic serine protease (proteolytic) enzyme with specificity for peptide bonds involving the carboxyl group of the basic amino acids, Arginine and Lysine. Trypsin often contains a crude mixture of lipases, nucleases, polysaccharides and proteases extracted from Porcine pancreas.

Most cell cultures grow as a single thickness cell layer or sheet attached to a substrate known as a monolayer. When subculturing adherent cells, these intercellular and cell-to-substrate links or connections must be gently dissociated. Proteolytic enzymes such as Trypsin (i.e. a serine peptidase), breaks or gently separates these bonds by creating a single-cell suspension from which new subcultures are realized.

Trypsin solutions are widely utilized as cell dissociation reagents for continuous cell culture of adherent growing cells. Trypsin proteolysis or trypsinization is a process in which proteins have been digested or treated with Trypsin and are thus said to be trypsinized. Biological Industries' Trypsin is designed not only to gently dissociate cells from almost any support substrates but also as well as from each other in order to actualize cell manipulation techniques in addition for other studies that require intact cell-surface proteins. Trypsin, as a solution, is available in a varied array of formulations with or without EDTA. EDTA is a chelator that binds calcium and magnesium ions that may otherwise inhibit the Trypsin activity which then hydrolyzes and gains access to the intercellular bonds being cell-cell and/or cell-substrate bonds. Crude Trypsin is often the subculturing agent of choice for cell dissociation/disaggregation of adherent cells although the treatment may be cytotoxic if prolonged. Over-trypsinization is a common cause of subculture problems.

The use of Crude Trypsin often involves multiple changes and the variability among lots can dramatically influence the effectiveness of the dissociation process. Regarding the use of Crude Trypsin, some important facts must be noted: 

v Cells must NEVER remain in the Crude Trypsin for longer than 3-5 minutes as they may be seriously damaged in the process (i.e. damage to the intracellular proteins).

v Cells should NEVER be left without a fluid layer

v Do not permit prolonged growth on culture ware as the cells will be very difficult to remove(i.e. after 5-7 days)

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