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专业: 病原细菌与放线菌生物学

[求助] 药物渗透性小鼠模型

各位高手们，我要做下面的实验，下面两段是我摘的两篇文献上的片段
2.1 药物渗透模型
首先对动物背部进行脱毛，然后麻醉，一般采用戊巴比妥麻醉动物（30mg/ kg），麻醉成功后用砂纸对动物背部进行打磨，用合适大小的小桶粘附与动物背部打磨部位，接着在小筒内进行给药。小筒一般直径1～2cm即可，要注意打磨程度。一般可在给药后1h、2h、4h进行检测[3]。药物可通过高效液相色谱法（HPLC）和免疫印迹法（western blot）进行检测。可观察得到药物透皮能力，监测药物的有效作用。此模型适用于大鼠、小鼠等动物。

The mice were anesthetized via intramuscular injections of Zoletil 50 at a dosage of 50 mg/kg of body weight. In order to assess the skin permeability of HG1, the shaved back skin of each experimental mouse was scratched with No. 600 sandpaper. A plastic cylinder with a diameter of 1.65 cm was affixed to the abrasion site using cyanoacrylate adhesive. A 200 ul volume of 1% (wt/vol) HG1 solution (10 mM sodium phosphate [NaP] buffer [pH 7.4] [10%], polyethylene glycol 400 [45%], glycerol [45%]) was poured into the cylinder chamber. The cylinder was removed at a predetermined time, after which the skin was washed extensively with NaP buffer (pH 7.4) to completely remove any HG1 that might remain on the surface of the skin. An area of skin (1-by-1 cm) was then excised and laid out on a slide glass. The skin samples were then frozen at - 80°C for 1 h. A constant volume of tissue was obtained from each of the frozen skin samples by the use of a biopsy punch with a diameter of 8 mm. The isolated tissues were homogenized in liquid nitrogen, and the proteins were extracted from the homogenates via overnight shaking in 0.5 ml of 5% acetic acid. After brief centrifugation, 0.1 ml of the supernatant was subjected to C18 reversed-phase high-performance liquid chromatography (HPLC)

我按照这篇英文文献上的步骤这么做，但是在离心取上清的时候遇到问题了，因为在跑HPLC之前样品要过0.22um滤膜，但是吸上来的上清是胶状的，根本过不了滤膜，所以请各位帮我分析一下，谢谢了。

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1楼2015-03-29 13:09:16

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