Pronase

Autor: Byron Th. Aftonomos, John F. Foley
Rok vydání: 1973
Předmět:
DOI: 10.1016/b978-0-12-427150-0.50051-6
Popis: Publisher Summary This chapter focuses on cell harvesting with pronase. Pronase is the trade name of a crude preparation of proteolytic enzyme(s) obtained from a broth culture of Streptomyces griseus, used in the manufacture of streptomycin. It has been found to more rapidly and efficiently disperse monolayer fibroblastic cell lines than any other available enzymatic method. It also disperses epithelial monolayers although it seldom provides single cell dispersion suitable for cloning experiments. Five to seven components have been demonstrated chromatographically in pronase, the major one being a neutral protease. Pronase can hydrolyze a large number of protein peptide bonds with the subsequent liberation of 70 to 90% of free amino acids, in comparison with acid hydrolysis. The enzyme exhibits maximum activity at pH 7 to 8 and high stability at pH 6 to 9 at room temperature. The stability markedly declines below pH 4 and above pH 10. The chapter explains that pronase is the single best proteolytic agent at present available for the dispersion of fibroblastic cell lines. It is superior to trypsin not only in separating monolayers from their substrate but also in producing single cell suspensions.
Databáze: OpenAIRE