| 查看: 2707 | 回复: 7 | ||
z631535213木虫 (正式写手)
|
[求助]
关于细胞裂解的一点问题 已有3人参与
|
|
准备自己配点裂解液用用，看了些东西，发现一些裂解液中提到有效裂解成分，我目前只看到三个，Triton X-100，NP40，SDS。 想问下对这方面有了解的虫友 1.曲通，NP40，SDS一般多少浓度可有效裂解细胞。主要是用来裂解细胞提取DNA来进行后面的实验 2.曲通，NP40，SDS对细胞的裂解作用能具体讲一讲么。 恩，大概就这些。拜谢  |
回复此楼» 猜你喜欢
筑牢营养安全线：以精准检测，护健康基石
已经有0人回复
-
推荐一些20种氨基酸检测的实际应用案例
已经有0人回复
-
化学工程及工业化学论文润色/翻译怎么收费?
已经有55人回复
-
不合理蛙科研实验中的趣事：实验器材的 “乌龙”
已经有0人回复
-
不合理蛙科研实验中的趣事：和实验材料的 “斗智斗勇”
已经有0人回复
-
蛋白质检测：精准分析，解锁生物分子的密码
已经有0人回复
-
不合理蛙科研实验之小鼠实验：严谨设计，解析生命机制的重要载体
已经有0人回复
-
不合理蛙科研实验之重金属检测：精准筛查，守护健康与环境的防线
已经有0人回复
-
不合理蛙科研实验之“蛙测重金属我背锅三千”
已经有0人回复
-
不合理蛙科研实验之“鼠逃三次我发三篇SCI”
已经有0人回复
» 本主题相关价值贴推荐，对您同样有帮助:
- 有木有谁建立细胞株的呀，求经验
已经有16人回复
- 动物组织研磨和裂解问题
已经有13人回复
- 关于293T细胞的问题，急求助
已经有43人回复
- 170kDa膜蛋白MRP1跑WB遇到问题
已经有39人回复
- ALP活性检测中细胞裂解问题
已经有10人回复
- 关于悬浮细胞MTT的一点疑问，求解答！
已经有11人回复
- 求助：在不破坏组蛋白结构的前提下，应该采用什么样的细胞裂解方法？
已经有12人回复
- 求助怎样使细菌细胞裂解但胞内的蛋白（例如酶）不失活？
已经有9人回复
- 细胞裂解提取细胞内的总蛋白，需要使用什么试剂盒，求推荐
已经有14人回复
- 细胞裂解缓冲液的配制？求助各位大虾
已经有3人回复
- 关于THP-1细胞的培养，分化问题
已经有12人回复
- 关于源内裂解的问题
已经有3人回复
- 关于质谱裂解规律的问题
已经有6人回复
- 求助裂解细胞又不降低酶活
已经有5人回复
- 【求助/交流】ALP的定量检测一些问题
已经有6人回复
- 【求助/交流】请各位帮帮忙~~细胞裂解液如何保存，能保存多长时间？
已经有8人回复
- 【求助/交流】为什么条带很细啊？郁闷呢！
已经有9人回复
- 【求助/交流】细胞裂解的问题
已经有19人回复
- 【求助】关于裂解气质联用分析的问题
已经有4人回复
z631535213
木虫 (正式写手)
- 应助: 9 (幼儿园)
- 金币: 2458.4
- 红花: 3
- 帖子: 479
- 在线: 113.5小时
- 虫号: 2756319
- 注册: 2013-10-27
- 性别: GG
- 专业: 基因组学
2楼2014-10-17 08:28:43
jurkat.1640
至尊木虫 (文坛精英)
- MolEPI: 1
- 应助: 905 (博后)
- 金币: 16549.9
- 散金: 1654
- 红花: 120
- 沙发: 41
- 帖子: 14459
- 在线: 2171.4小时
- 虫号: 514340
- 注册: 2008-02-28
- 性别: GG
- 专业: 病毒学
3楼2014-10-17 08:58:00
z631535213
木虫 (正式写手)
- 应助: 9 (幼儿园)
- 金币: 2458.4
- 红花: 3
- 帖子: 479
- 在线: 113.5小时
- 虫号: 2756319
- 注册: 2013-10-27
- 性别: GG
- 专业: 基因组学
4楼2014-10-17 10:31:23
5楼2014-10-22 21:34:59
6楼2014-10-23 10:03:54
【答案】应助回帖
|
Detergents are amphipathic molecules, meaning they contain both a nonpolar "tail" having aliphatic or aromatic character and a polar "head". Ionic character of the polar head group forms the basis for broad classification of detergents; they may be ionic (charged, either anionic or cationic), nonionic (uncharged) or zwitterionic (having both positively and negatively charged groups but with a net charge of zero). Detergents can be denaturing or non-denaturing with respect to protein structure. Denaturing detergents can be anionic such as sodium dodecyl sulfate (SDS) or cationic such as ethyl trimethyl ammonium bromide. These detergents totally disrupt membranes and denature proteins by breaking protein:protein interaction. Non-denaturing detergents can be divided into nonionic detergents such as Triton* X-100, bile salts such as cholate and zwitterionic detergents such as CHAPS. A major factor determining the behavior and interaction of molecules in biological samples is their hydrophilicity or hydrophobicity. Most proteins and other molecules with charged or polar functional groups are soluble (or miscible) in water because they participate in the highly ordered, hydrogen-bonded intermolecular structure of water. Some other proteins (or at least parts of proteins), as well as fats and lipids, lack polar or charged functional groups; consequently, they are excluded from the ordered interaction of water with other polar molecules and tend to associate together in structures having minimal surface area contact with the polar environment. This association of nonpolar molecules in aqueous solutions is commonly called hydrophobic attraction. Generally, moderate concentrations of mild (i.e., nonionic) detergents compromise the integrity of cell membranes, thereby facilitating lysis of cells and extraction of soluble protein, often in native form. Using certain buffer conditions, various detergents effectively penetrate between the membrane bilayers at concentrations sufficient to form mixed micelles with isolated phospholipids and membrane proteins. Detergent monomers solubilize membrane proteins by partitioning into the membrane bilayer. With increasing amounts of detergents, membranes undergo various stages of solubilization. The initial stage is lysis or rupture of the membrane. At detergent:membrane lipid molar ratios of 0.1:1 through 1:1 the lipid bilayer usually remains intact but selective extraction of some membrane proteins occurs. Increasing the ratio to 2:1, solubilization of the membrane occurs resulting in mixed micelles. These include phospholipid-detergent micelles, detergent-protein micelles, and lipid-detergent-protein micelles. At a ratio of 10:1, all native membrane lipid:protein interactions are effectively exchanged for detergent:protein interactions |
7楼2014-10-23 10:18:14
z631535213
木虫 (正式写手)
- 应助: 9 (幼儿园)
- 金币: 2458.4
- 红花: 3
- 帖子: 479
- 在线: 113.5小时
- 虫号: 2756319
- 注册: 2013-10-27
- 性别: GG
- 专业: 基因组学
8楼2014-10-23 12:43:54