| 查看: 667 | 回复: 3 | ||
musejing1银虫 (初入文坛)
|
[求助]
求翻译 已有1人参与
|
|
Here 5μl of each activated medium was mixed with 1 ml of 0.5 M ethanolamine–HCl solution (pH 7.5) for the purpose of blocking the NHS or aldehyde (CHO) group. The resultant mixture was then stirred at room temperature for 2 h. After removing the supernatant by centrifugation, the particle was washed twice with 1 ml of phosphate-buffered saline (PBS). In the case of NHS-activated media, the amount of immobilized NHS was calculated according to the absorbance of the supernatant at 280 nm. Human serum solution (1 ml, diluted 5with PBS) was applied to the medium, and the mixture was stirred at room temperature for 1 h. After that, the resultant mixture was centrifuged and the medium was washed seven times with 1 ml of PBS. Subsequently, 100μl of 0.2 M glycine–HCl (pH 2.5) was applied to the medium, and the mixture was stirred at room temperature for 0.5 h. The resultant mixture was centrifuged, and 50ll of supernatant, 50ll of 1.0 M Tris–HCl (pH8.3), and 100ll of Laemmli sample buffer (Bio-Rad Laboratories, Hercules, CA, USA) were mixed and heated in boiling water for 3 min (acid elution fraction). Furthermore, after washing the particle twice with 0.2 M glycine–HCl (pH 2.5), 100ll of Laemmli sample buffer was applied to the particle and heated in boiling water for 3 min. After that, the resultant mixture was centrifuged and the supernatant was obtained (sodium dodecyl sulfate [SDS] elution fraction). These elution fractions were applied to a 5–15% acrylamide gel (Bio-Rad Laboratories), and SDS–polyacrylamide gel electrophoresis (PAGE) was performed. The gel was stained with a silver staining kit (Silver Stain II Kit, Wako Pure Chemical Industries), and the level of nonspecifically adsorbed proteins was confirmed。 |
回复此楼» 猜你喜欢
有多少人是今天查系统知道结果的？
已经有19人回复
-
29号明天会评吗
已经有4人回复
-
国社科又开始会评了，不知道这次命运如何
已经有10人回复
-
梦想
已经有4人回复
-
中青基了要发朋友圈吗？
已经有6人回复
-
基金不中，共勉
已经有12人回复
-
能否申诉？
已经有7人回复
-
面上意见出来了
已经有19人回复
-
为什么到现在没收到通知？
已经有5人回复
-
有没有仍没收到信息的
已经有8人回复
1016828262
铁虫 (小有名气)
- 应助: 7 (幼儿园)
- 金币: 125.3
- 散金: 15
- 红花: 3
- 帖子: 59
- 在线: 13.4小时
- 虫号: 2371235
- 注册: 2013-03-23
- 专业: 微生物生理与生物化学
2楼2014-03-31 20:26:26
1016828262
铁虫 (小有名气)
- 应助: 7 (幼儿园)
- 金币: 125.3
- 散金: 15
- 红花: 3
- 帖子: 59
- 在线: 13.4小时
- 虫号: 2371235
- 注册: 2013-03-23
- 专业: 微生物生理与生物化学
3楼2014-03-31 20:28:09
musejing1
银虫 (初入文坛)
- 应助: 0 (幼儿园)
- 金币: 465.5
- 散金: 5
- 帖子: 9
- 在线: 6.6小时
- 虫号: 2826530
- 注册: 2013-11-25
- 性别: MM
- 专业: 无机合成和制备化学
4楼2014-04-03 14:01:33
10