Isolation of monkey aortic lysosomes using Percoll density gradient.
A novel technique involving the Percoll density gradient and 0.01M phosphate buffer has been employed for the first time on aortic tissue for isolation of lysosomes. The purity of the lysosomes has been established by marker-enzymes, acid phosphatase and N-acetyl-beta-D-glucosaminidase and latent activities of lysosomal hydrolases. The heavier fraction (density 1.08) obtained after Percoll density gradient centrifugation showed high specific activities of lysosomal hydrolases and these enzymes were markedly latent. Moreover this heavier (lysosome rich) fraction has been noted to be free of other sub-cellular contaminants.
Dhaunsi
G S
GS
Department of Experimental Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Majumdar
S
S
Chakravarti
R N
RN
D000118:Acetylglucosaminidase / Q000302:isolation & purification; D000135:Acid Phosphatase / Q000302:isolation & purification; D000818:Animals; D001011:Aorta / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D006867:Hydrolases / Q000302:isolation & purification; D008247:Lysosomes / Q000201:enzymology; D008253:Macaca mulatta; D008297:Male; D008722:Methods
D006867:Hydrolases; D000135:Acid Phosphatase; D000118:Acetylglucosaminidase