Andersen K J, Schjønsby H, Skagen D W
Scand J Gastroenterol. 1983 Mar;18(2):241-9. doi: 10.3109/00365528309181590.
To define reproducible conditions for the homogenization of small-intestinal biopsy samples, tissue homogenization has been studied by the use of three different homogenizers. Tissue samples of increasing wet weights (0.5-10.8 mg) were homogenized in a fixed volume (1 ml) before DNA and protein were determined. The DNA to protein ratio was calculated for all wet weights and used as a measure for reproducible homogenization. The minimum tissue wet weight needed for analysis (2 mg) was determined from the values obtained for the DNA to protein ratio. Highly sensitive techniques are described in detail for the assay of brush border (maltase, lactase, sucrase, neutral alpha-glucosidase, alkaline phosphatase, gamma-glutamyl transferase, leucyl-beta-naphthylamidase), basolateral membrane (5'-nucleotidase), and mitochondrial (succinate dehydrogenase) marker enzymes and for four acid hydrolases (acid phosphatase, acid beta-D-galactosidase, N-acetyl-beta-D-glucosaminidase, acid diesterase) in human and rat jejunal mucosa. Linear kinetics have been established for all enzyme assays. The optimal dilution of tissue homogenate for the assay of the various enzymes has been determined to enable the determination of a maximum number of enzymes in each homogenate. The range of enzyme activities in samples of human and rat jejunal mucosa has been determined.
为确定小肠活检样本匀浆的可重复条件，已使用三种不同的匀浆器对组织匀浆进行了研究。在测定DNA和蛋白质之前，将湿重不断增加（0.5 - 10.8毫克）的组织样本在固定体积（1毫升）中匀浆。计算所有湿重下的DNA与蛋白质比率，并将其用作可重复匀浆的指标。根据DNA与蛋白质比率获得的值确定分析所需的最小组织湿重（2毫克）。详细描述了用于检测人及大鼠空肠黏膜刷状缘（麦芽糖酶、乳糖酶、蔗糖酶、中性α - 葡糖苷酶、碱性磷酸酶、γ - 谷氨酰转移酶、亮氨酰 - β - 萘酰胺酶）、基底外侧膜（5'-核苷酸酶）和线粒体（琥珀酸脱氢酶）标记酶以及四种酸性水解酶（酸性磷酸酶、酸性β - D - 半乳糖苷酶、N - 乙酰 - β - D - 葡糖胺酶、酸性二酯酶）的高灵敏度技术。已为所有酶测定建立了线性动力学。已确定用于各种酶测定的组织匀浆的最佳稀释度，以便能够在每个匀浆中测定最多数量的酶。已确定人及大鼠空肠黏膜样本中的酶活性范围。
