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243562850新虫 (初入文坛)
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[求助]
合成的抗体-碳纳米管作用在细胞怎样灭菌?
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大家好,我现在在按着文献的步骤往碳纳米管上连接单克隆抗体,步骤是下面这样的: The Preparation of anti-P-gp Functionalized Carbon Nanotubes (Ap-SWNTs). The AP-SWNTs were prepared by covalently bonding the anti-P-gp antibody on the o-SWNTs via the diimide-activated amidation process as described in the literature.56 The scheme for this preparation is presented in Figure 1. Briefly, 5 mg of the water-soluble o-SWNTs, 30 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), and 300 mg of N-hydroxysuccinimide (NHS) were added in a 10 mL aqueous buffer solution of 2-(4-morpholino)ethanesulfonic acid (MES) (50 mM, pH 6.0), mixed, and shaken in a reciprocating shaker at room temperature for 2 h. After that, the resultant SWNTs were filtered through a 0.45 m filter, fully rinsed with 50 mM MES buffer, and redissolved in a 10 mL MES buffer solution (50 mM, pH 6.0). Subsequently, 1 mL of anti-P-gp solution (0.025 mg/mL) was added, and the mixture was shaken at room temperature for 2 h. This was followed by a filtration through 0.45 μm filter and a consecutive rinse with 2 mL of NaCl solution (0.1 M) until there was no antibody detected in the filtrate by a Hitachi F-2500 fluorescence spectrophotometer (FS) (Pleasanton, CA,USA) with excitation at 488 and emission at 533 nm. Finally, the resulting Ap-SWNTs were collected, lyophilized to dryness, and stored at 4 °C before use (TEM image of Ap-SWNTs shown in Supporting Information, Figure S-1). 我想请教大家一个问题,就是做在细胞上的时候为防止细胞污染,抗体-碳纳米管要怎样处理才可以呢?谢谢! |
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