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一种使用十二烷基硫酸钠使肝甘油三酯脂肪酶失活来测量肝素后血浆中脂蛋白脂肪酶的新方法。

A new method for the measurement of lipoprotein lipase in postheparin plasma using sodium dodecyl sulfate for the inactivation of hepatic triglyceride lipase.

作者信息

Baginsky M L, Brown W V

出版信息

J Lipid Res. 1979 May;20(4):548-56.

PMID:458271
Abstract

Lipoprotein lipase (LPL) and hepatic triglyceride lipase (H-TGL) are lipolytic activities found in postheparin plasma. A simple and precise method for the direct determination of LPL in postheparin plasma is described. Pre-incubations of this plasma (45--60 min at 26 degrees C) with sodium dodecyl sulfate (35--50 mM) in 0.2 M Tris-HCl buffer, pH 8.2, results in the inactivation of H-TGL, while leaving LPL fully active. Direct determination of H-TGL is done in a separate aliquot of the same postheparin plasma sample using previously reported assay conditons that do not measure LPL. The sodium dodecyl sulfate-resistant lipolytic activity has the characteristics of LPL as judged by a) its activation by serum and by apolipoprotein C-II; b) its inactivation (over 90%) by 0.75 M NaCl; and c) its inactivation by a specific antiserum. No sodium dodecyl sulfate-resistant activity was found in postheparin plasma from a patient with LPL deficiency (primary type I hyperlipoproteinemia). An excellent correlation of values was obtained (r = 0.99) for 30 samples assayed after sodium dodecyl sulfate treatment and after immuno-inactivation of H-TGL. The intra-assay coefficient of variation was +/- 11% and 4% before and after normalization of values, respectively.

摘要

脂蛋白脂肪酶(LPL)和肝甘油三酯脂肪酶(H-TGL)是在肝素后血浆中发现的脂解活性。本文描述了一种直接测定肝素后血浆中LPL的简单而精确的方法。将该血浆在0.2M Tris-HCl缓冲液(pH 8.2)中与十二烷基硫酸钠(35-50 mM)预孵育(26℃下45-60分钟),可使H-TGL失活,而LPL仍保持完全活性。使用先前报道的不检测LPL的测定条件,在同一肝素后血浆样品的另一等分试样中直接测定H-TGL。通过以下几点判断,耐十二烷基硫酸钠的脂解活性具有LPL的特征:a)其被血清和载脂蛋白C-II激活;b)其被0.75M NaCl灭活(超过90%);c)其被特异性抗血清灭活。在患有LPL缺乏症(原发性I型高脂蛋白血症)的患者的肝素后血浆中未发现耐十二烷基硫酸钠的活性。对30个样品在十二烷基硫酸钠处理后和H-TGL免疫灭活后进行测定,得到了极好的相关性(r = 0.99)。在数值标准化前后,测定内变异系数分别为+/- 11%和4%。

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