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[求助]
四环庚烷连接反应文献中的图解，求高人指点啊 已有1人参与
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Figure 2. Ni bis(dithiolene) reagents selectively label quadricyclane-modified BSA. (A) Modification of BSA’s solvent-exposed lysine residues with compound 16 and subsequent labeling with biotinylated Ni bis(dithiolene) reagent 15. (B, C) Ni bis (dithiolene) 15 displays (B) time- and (C) dose-dependent labeling of QC-BSA. BSA ( ) or QC-BSA (+) (5 μg) was treated with (B) 50 μM 15 for various amounts of time or (C) various concentrations of 15 for 30 min. The reactions were quenched with 1 and 5, and the presence of product was detected by Western blot using α-biotin -HRP. (D) Reaction of 15 with QC-BSA can be prevented by pretreatment with tetr- asulfonated Ni bis(dithiolene) 17. BSA ( ) or QC-BSA (+) (5 μg) was treated with var- ious amounts of 17 for 30 min followed by 50 μM 15 for 30 min. The react- ions were quenched with 1 and 5, and the samples were analyzed by Western blot probing with α-biotin-HRP. (E) QC-BSA can be selectively labeled in a mixture of proteins. To 25 μg of lysate in the presence of 1 mM K3Fe(CN)6 was added no BSA or reagent (lane 1), BSA (1.5 μg) and 50 μM 15 (lane 2), and QC-BSA (1.5 μg) and 50 μM 15 (lane 3). After 30 min, the reactions were quenched with 1 and 5 and analyzed by Western blot. The BSA monomer and dimer bands are visible in the QC-BSA-treated sample. The bands denoted with asterisks represent a biotinyla- ted E. coli protein. In (B E), equal protein loading was verified by Ponceau stain. 专业知识匮乏，实验现象图的解释完全看不懂啊，太多专有名词，请专业人士翻译一下呗  1111.png |
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G欧尼~(草也花开代发): 金币+8, 谢谢您的认真回复，欢迎多来专外版交流 2016-07-21 07:34:21
G欧尼~(草也花开代发): 金币+8, 谢谢您的认真回复，欢迎多来专外版交流 2016-07-21 07:34:21
| Ni二（二硫烯）试剂选择性标记四环庚烷修饰的BSA。（A）对BSA暴露在溶剂中的赖氨酸残基用化合物16修饰，进而用生物酰化的Ni二（二硫烯）试剂15进行标记。（B,C）Ni二（二硫烯）试剂15对QC-BSA的标记表现出受时间（B）和剂量（C）的影响。BSA或QC-BSA(5ug)和50uM的试剂15反应不同的时间（B）或与不同浓度的15反应30min(C).反应液与1和5混合，产物通过Western斑点印迹实验使用α-生物素HRP进行检测。（D）15和QC-BSA的反应可以通过用四磺酰化的镍二（二硫烯）17预处理来避免.BSA或QC-BSA5ug和不同剂量的17反应30min然后与50uM的15反应30min。反应液与1和5混合，产物通过Western斑点印迹实验使用α-生物素HRP进行检测。（E）QC-BSA在蛋白质混合物中能被选择性标记。向加入1mM的K3Fe(CN)6的25ug的溶解产物中加入BSA(0ug)和0uM的15（路线1）BSA(1.5ug)和50uM15（路线2）和QC-BSA(1.5ug)和50uM的15（路线3）。30min后，反应液与1和5混合，产物通过Western斑点印迹实验使用α-生物素HRP进行检测。BSA单体和二聚物的带在QC-BSA处理的样品中是可见的。星号标记的带代表生物酰化的E，大肠杆菌蛋白质。在（BE）中，相等的蛋白质量是通过丽春江染色来区分的。 |
2楼2016-07-05 09:24:59
