[Effective extraction of lysosomal enzymes with digitonin].
A method for rapid and effective extraction of rat liver lysosomal enzymes has been elaborated. It includes isolation of lysosomal-mitochondrial fraction by means of differential centrifugation, selective destruction of the lysosomal membrane by digitonin and centrifugal obtaining of the lysosomal matrix. Total labilization of the lysosomal membrane is achieved at 0.3 mM of the detergent. The maximal enrichment of an extract by lysosomal enzymes is observed in the range of 0.3-0.4 mM of digitonin. The level of lysosomal enzyme purification is 30.7 for cysteine cathepsins B, L, H, 24.9- for beta-galactosidase, 14.1- for acid phosphatase. The method gives high yield of lysosomal enzymes (40-80%).
Pupyshev
A B
AB
Korolenko
T A
TA
D000818:Animals; D002458:Cell Fractionation; D004072:Digitonin / Q000494:pharmacology*; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains
D004072:Digitonin
Effektivnaia ékstraktsiia lisosomal'nykh fermentov s pomoshch'iu digitonina.