Dhaunsi G S, Majumdar S, Chakravarti R N
Department of Experimental Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Biochem Int. 1987 May;14(5):787-95.
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.
一种涉及Percoll密度梯度和0.01M磷酸盐缓冲液的新技术首次应用于主动脉组织以分离溶酶体。通过标记酶、酸性磷酸酶和N-乙酰-β-D-氨基葡萄糖苷酶以及溶酶体水解酶的潜在活性来确定溶酶体的纯度。Percoll密度梯度离心后获得的较重部分(密度1.08)显示出溶酶体水解酶的高比活性,并且这些酶明显处于潜在状态。此外,已注意到这个较重的(富含溶酶体的)部分不含其他亚细胞污染物。