Analysis of picogram quantities of protein in subnanoliter-size samples.
Analysis of picogram quantities of protein in subnanoliter-size samples.
复制标题
分析亚纳升样品中皮克量的蛋白质。
DOI：
10.1016/0003-2697(84)90535-9
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发表时间：
1984
影响因子：
2.9
通讯作者：
Roman,RJ
中科院分区：
文献类型：
--
作者：
Zawieja,D;Barber,BJ;Roman,RJ
测量亚纳升体积蛋白质浓度的能力将有助于许多生物学研究。开发了一种用于测量纳升样品中纳克蛋白质含量的微量测定法和一种用于测量皮升样品中皮克含量的超微量测定法来测量淋巴蛋白质浓度。使用微量移液技术将样品溶液的等分试样与邻苯二甲醛巯基乙醇试剂混合。使用改进的 Leitz MPV-1 显微镜作为显微荧光计测量反应产物荧光。荧光随白蛋白浓度在 1 至 8 g/100 ml 之间呈线性变化。测量 0.0、1.0、2.0 和 4.0 g/100 ml ml 的白蛋白标准品的典型微量测定得出 y = 207x + 60 (r = 0.99) 的线性回归。最低可检测蛋白质浓度为 0.125 g/100 ml。白蛋白标准品的 SE 范围为 0.02 至 0.17 g/100 ml。测量类似标准的超微量测定产生 y = 1180x + 109 (r = 0.96) 的线性回归。最低可检测蛋白质浓度为 0.028 g/100 ml。标准品的 SE 范围为 0.01 至 0.32 g/100 ml。
The ability to measure protein concentration in subnanoliter volumes would be helpful in many biological studies. A microassay for measuring nanogram protein quantities in nanolitersize samples and an ultramicroassay for measuring picogram quantities in picoliter samples were developed to measure lymphatic protein concentration. Aliquots of a sample solution were mixed with an o-phthalaldehyde mercaptoethanol reagent using micropipetting techniques. Reaction product fluorescence was measured using a modified Leitz MPV-1 microscope as a microfluorometer. Fluorescence varied linearly with albumin concentrations between 1 and 8 g/100 ml. A typical microassay measuring albumin standards at 0.0, 1.0, 2.0, and 4.0 g/100 ml ml yielded a linear regression of y = 207x + 60 (r = 0.99). Minimum detectable protein concentration was 0.125 g/100 ml. The SE for the albumin standards varied from 0.02 to 0.17 g/100 ml. An ultramicroassay measuring similar standards yielded a linear regression of y = 1180x + 109 (r = 0.96). Minimum detectable protein concentration was 0.028 g/100 ml. The SE for the standards varied from 0.01 to 0.32 g/100 ml.