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Gel Electrophoresis Method to Measure the Concentration of Single-Walled Carbon Nanotubes Extracted from Biological Tissue

机译:凝胶电泳法测量从生物组织中提取的单壁碳纳米管的浓度

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摘要

A rapid and sensitive method to detect single-walled carbon nanotubes (SWNTs) in biological samples is presented. The method uses polyacrylamide gel electrophoresis (PAGE) followed by quantification of SWNT bands. SWNTs dispersed in bovine serum albumin (BSA) were used to develop the method. When BSA- SWNT dispersions were subjected to sodium dodecyl sulfate (SDS)-PAGE, BSA passed through the stacking gel, entered the resolving gel, and migrated toward the anode as expected. The SWNTs, however, accumulated in a sharp band at the interface between the loading well and the stacking gel. The intensities from digitized images of these bands were proportional to the amount of SWNTs loaded onto the gel with a detection limit of 5 ng of SWNTs. To test the method, normal rat kidney (NRK) cells in culture were allowed to take up SWNTs upon exposure to medium containing various concentrations of BSA-SWNTs for different times and temperatures. The SDS-PAGE analyses of cell lysate samples suggest that BSA-SWNTs enter NRK cells by fluid-phase endocytosis at a rate of 30 fg/day/cell upon exposure to medium containing 98 (mu)g/mL SWNTs.
机译:提出了一种快速灵敏的方法来检测生物样品中的单壁碳纳米管(SWNTs)。该方法使用聚丙烯酰胺凝胶电泳(PAGE),然后定量SWNT条带。分散在牛血清白蛋白(BSA)中的SWNT用于开发该方法。当对BSA-SWNT分散液进行十二烷基硫酸钠(SDS)-PAGE处理时,BSA穿过堆积凝胶,进入拆分凝胶,并按预期向阳极迁移。但是,单壁碳纳米管在装载井和堆积凝胶之间的界面处以尖锐的带状积累。这些条带的数字化图像的强度与加载到凝胶上的SWNT的数量成比例,检测极限为5 ng SWNT。为了测试该方法,在暴露于含有不同浓度的BSA-SWNT的培养基中,在不同的时间和温度下,允许培养的正常大鼠肾(NRK)细胞摄取SWNT。细胞裂解物样品的SDS-PAGE分析表明,当暴露于含有98μg/ mL SWNTs的培养基中时,BSA-SWNTs通过液相内吞作用以30 fg / day /细胞的速度进入NRK细胞。

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