| 查看: 313 | 回复: 1 | |||
[求助]
帮忙翻译
|
|
The oxidation was carried out in 23 mL of a phosphate buffer (100 mm) containing 2 mm EDTA, 3 mm Cys, 0.15 mm Cys2 and 6 m GunHCl with a peptide concentration of 100 mg/mL. The reaction mixture was dialysed (2 h each) against 1 L of the same buffer containing 4 m GunHCl (2 h), 2 m GunHCl (2 h) and the buffer without GunHCl (20 h). The fully oxidized product was then purified by HPLC (Vydac C18, 103250 mm, 300 A° , 5 mm, flow rate 3 mL/min; eluent A, 0.07% TFA; eluent B, 0.07% TFA in MeCN–H2O 80 : 20; gradient: 10–70% B in 60 min, UV detection at 215 nm). Purity was checked on a Nucleosil C18 PPN column as described above and by capillary electrophoresis (P/ACE System 2000, Beckman, Munich, Germany; fused silica capillary, 47 cm375 mm, uncoated; 0.1 m phosphate buffer containing 0.02% hydroxymethylpropylcellulose, pH 2.5; constant current 120 mA; UV detection at 200 nm). The molecular weight determined for LEAP-1/hepcidin by ESIMS was 2789.5 (Mr calc. 2789.3). Trypsin cleavage of oxidized b-defensins A sample of 300 mg of the purified defensins was dissolved in 500 mL of 100 mm Tris-HCl. A solution of 12.5 mg trypsin in 1 mm HCl (25 mL) was added. After 24 h at 368C, the reaction was stopped by separation of the mixture (100 mL) on an analytical Vydac C18 HPLC column. The tryptic fragments detected were collected and analysed by ESIMS, and the amino acid sequences of selected defensin fragments were determined by Edman degradation (494 Procise peptide sequencer, PE Biosystems, Weiterstadt, Germany). |
回复此楼» 猜你喜欢
情人节自我反思：在爱情中有过遗憾吗？
已经有6人回复
-
基金正文30页指的是报告正文还是整个申请书
已经有4人回复
-
今年春晚有几个节目很不错，点赞！
已经有6人回复
-
球磨粉体时遇到了大的问题，请指教！
已经有15人回复
-
过年走亲戚时感受到了所开私家车的鄙视链
已经有5人回复
-
江汉大学解明教授课题组招博士研究生/博士后
已经有3人回复
zerohead
禁虫 (职业作家)
- 翻译EPI: 466
- 应助: 1 (幼儿园)
- 贵宾: 0.301
- 金币: 8727.8
- 散金: 850
- 红花: 15
- 沙发: 10
- 帖子: 3768
- 在线: 471.7小时
- 虫号: 1041741
- 注册: 2010-06-14
- 性别: GG
- 专业: 逻辑学
2楼2010-07-14 09:48:42