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396101466金虫 (正式写手)
一段遗失的美好
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【求助/交流】有关免疫沉淀电泳的的操作 已有5人参与
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| 最近看了一篇文章，做了免疫沉淀分析，并作了电泳，但是不清楚电泳中的的成分是沉淀还是上清，想问各位高人，免疫沉淀电泳的具体操作是怎么样的！ |
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4楼2011-04-25 17:52:36
yabxxh
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2楼2011-03-23 11:05:22
396101466
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silicare(金币+3): 很好，鼓励交流 2011-04-25 18:23:23
silicare(金币+3): 很好，鼓励交流 2011-04-25 18:23:23
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我只想解释两个图，图的说明是Fig. 5. Immunoprecipitation using the D1 (A) and A3 (B) Fab fragments and cow's milk samples. The Fab fragments were immobilised on protein L beads and then incubated with themilk samples without orwith heat treatment (0,15, 30 or 60 min at 95°C). After washing steps, the samples were analysed on a 15% SDS-PAGE gel and stained with silver. The boxes indicate the protein bands that were cut out for MS-analysis. Arrows indicate the Fab fragments. Ctr=protein L beads without the immobilized Fab incubated with 0′ (1) and 60′ (2) heated BLG sample. As a control, 1 µg of native BLG (Sigma) and D1/A3 Fab were used. 其操作内容是： Pasteurized, skimmed cow's milk was centrifuged (20 000×g, 2 min, 4 °C) and the supernatant was diluted 1:4 in PBS and used for immunoprecipitation. In the immunoprecipitation assay, both heat-treated (15, 30,60 min at 95 °C) and native cow's milk samples were used. The BLG-specific Fab fragments D1 and A3 were immobilized onto protein L beads (CBind™L, Fluka) according to the manufacturer's instructions (100-150 µg protein/25 µl beads). Then the milk samples were incubated with protein L-Fab beads for 1 h at room temperature. After the washing step with PBS-0.05% Tween 20, elution was performed by adding first 25 µl and then 50 µl of 0.1 M glycine-HCl (pH 2). The eluted samples were separated on a 15% SDS-PAGE gel and stained with silver. 我想问的是图中的条带怎么解释呀？谢谢啦！   |
3楼2011-03-23 15:22:59
ujsyangyong
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5楼2011-04-25 22:10:28