| 查看: 536 | 回复: 3 | |||
| 当前主题已经存档。 | |||
| 【有奖交流】积极回复本帖子，参与交流，就有机会分得作者 arniekyo 的 4 个金币 | |||
| 当前只显示满足指定条件的回帖，点击这里查看本话题的所有回帖 | |||
[交流]
【求助/交流】蛋白纯化 已有2人参与
|
|||
| 我现在要提取原核表达的蛋白，不知道相关的试剂上样buffer，漂洗buffer，洗脱buffer怎么样配制，是His标签的包涵体，请教高手指点 |
回复此楼» 猜你喜欢
含氟糖皮质激素的外用抗炎利器醋酸氟轻松
已经有0人回复
-
阿莫奈韦（Amenamevir）：解旋酶-引物酶靶向分子的结构特征与合成路径全解析
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133人回复
-
利瑞鲁单抗：靶向KIR的NK细胞检查点工具抗体，结构基础与科研应用全解析
已经有0人回复
-
Wnt-C59：PORCN靶向化学探针，Wnt信号通路阻断机制与科研应用全解析
已经有0人回复
-
Fexaramine：肠道限制性FXR化学探针，结构特征与信号通路研究全解析
已经有0人回复
-
NVS-STG2：靶向STING跨膜域的分子胶类激动剂，结构特征与寡聚化机制研究全解析
已经有0人回复
-
DOTA-GPC3肽（CAS 3066439-94-6）：双功能分子探针前体的结构特征与标记应用解析
已经有0人回复
-
科研用 TLR7/8 双激动剂怎么选？现货与质控要点
已经有0人回复
-
SEL120-34A HCl：CDK8/CDK19高选择性激酶抑制剂
已经有0人回复
» 本主题相关商家推荐: (我也要在这里推广)
jingcp200320
银虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 524.5
- 红花: 2
- 帖子: 218
- 在线: 12.6小时
- 虫号: 958529
- 注册: 2010-03-01
- 性别: GG
- 专业: 分子生物学
★ ★ ★
scelab(金币+3):热心虫友，欢迎多来交流~~~:tiger06::tiger28::tiger23: 2010-04-16 22:00
arniekyo(金币+1): 2010-04-17 19:42
arniekyo(金币+1): 2010-04-18 14:57
arniekyo(金币+1): 2010-06-18 09:34:05
scelab(金币+3):热心虫友，欢迎多来交流~~~:tiger06::tiger28::tiger23: 2010-04-16 22:00
arniekyo(金币+1): 2010-04-17 19:42
arniekyo(金币+1): 2010-04-18 14:57
arniekyo(金币+1): 2010-06-18 09:34:05
|
Recipes Buffer Stock Solutions (10X) To prepare the buffer solutions described below, you need to prepare sodium phosphate stock solutions: Stock Solution A (10X) 200 mM sodium phosphate, monobasic (NaH2PO4) 5 M NaCl Dissolve 27.6 g sodium phosphate, monobasic (NaH2PO4) and 292.9 g NaCl in 900 ml of deionized water. Mix well and adjust the volume to 1 L with deionized water. Store solution at room temperature. Stock Solution B (10X) 200 mM sodium phosphate, dibasic (Na2HPO4) 5 M NaCl Dissolve 28.4 g sodium phosphate, dibasic (Na2HPO4) and 292.9 g of NaCl in 900 ml of deionized water. Mix well and adjust the volume to 1 L with deionized water. Store solution at room temperature. 5X Native Purification Buffer 250 mM NaH2PO4, pH 8.0 2.5 M NaCl Prepare 200 ml solution as follows: 1. To 180 ml deionized water, add Sodium phosphate, monobasic 7 g NaCl 29.2 g 2. Mix well and adjust the pH with NaOH to pH 8.0. 3. Bring the final volume to 200 ml with water. 4. Store buffer at room temperature. 3 M Imidazole pH 6.0 3 M Imidazole 500 mM NaCl 20 mM Sodium Phosphate Buffer, pH 6.0 Prepare 100 ml solution as follows: 2. To 90 ml deionized water, add Imidazole 20.6 g Stock Solution A (10X) 8.77 ml Stock Solution B (10X) 1.23 ml 3. Mix well and adjust the pH to 6.0 with HCl or NaOH as necessary. 4. Bring the final volume to 100 ml with water. If the solution forms a precipitate, heat solution until the precipitate dissolves. 5. Store buffer at room temperature. Continued on next page 18 Recipes, Continued Guanidinium Lysis Buffer 6 M Guanidine Hydrochloride 20 mM Sodium Phosphate, pH 7.8 500 mM NaCl Prepare 100 ml solution as follows: 1. To 90 ml deionized water, add Stock Solution A (10X) 0.58 ml Stock Solution B (10X) 9.42 ml Guanidine Hydrochloride 57.3 g 2. Stir the solution until completely dissolved. Adjust the pH to 7.8 using 1 N NaOH or 1 N HCl. 3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 μm filter (autoclaving the solution will alter the pH of the buffer). 4. Store buffer at room temperature. Denaturing Binding Buffer 8 M Urea 20 mM Sodium Phosphate pH 7.8 500 mM NaCl Prepare 100 ml solution as follows: 1. To 90 ml deionized water, add Stock Solution A (10X) 0.58 ml Stock Solution B (10X) 9.42 ml Urea 48.1g 2. Stir the solution with gentle heating (50-60°C, do not overheat) until completely dissolved. When cooled to room temperature, adjust the pH to 7.8 using 1 N NaOH or 1 N HCl. 3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 μm filter (autoclaving the solution will alter the pH of the buffer). 4. Store buffer at room temperature. Continued on next page 19 Recipes, Continued Denaturing Wash Buffer 8 M Urea 20 mM Sodium Phosphate, pH 6.0 500 mM NaCl Prepare 100 ml solution as follows: 1. To 90 ml deionized water, add Stock Solution A (10X) 7.38 ml Stock Solution B (10X) 2.62 ml Urea 48.1g 2. Stir the solution with gentle heating (50-60°C, do not overheat) until completely dissolved. Adjust the pH to 6.0 using 1 N NaOH or 1 N HCl. 3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 μm filter (autoclaving the solution will alter the pH of the buffer). 4. Store buffer at room temperature. Denaturing Elution Buffer 8 M Urea 20 mM Sodium Phosphate, pH 4.0 500 mM NaCl Prepare 100 ml as follows: 1. To 90 ml deionized water, add Stock Solution A (10X) 10 ml Urea 48.1g 2. Stir the solution with gentle heating (50-60°C, do not overheat) until completely dissolved. Adjust the pH to 4.0 using 1 N NaOH or 1 N HCl. 3. Bring the volume to 100 ml and filter sterilize the buffer using a 0.45 μm filter (autoclaving the solution will alter the pH of the buffer). 4. Store buffer at room temperature. |
3楼2010-04-16 15:57:03
小白3521
金虫 (小有名气)
- 应助: 1 (幼儿园)
- 金币: 690.8
- 散金: 218
- 帖子: 251
- 在线: 73.8小时
- 虫号: 769519
- 注册: 2009-05-13
- 专业: 生物大分子结构与功能
2楼2010-04-16 14:50:31
jingcp200320
银虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 524.5
- 红花: 2
- 帖子: 218
- 在线: 12.6小时
- 虫号: 958529
- 注册: 2010-03-01
- 性别: GG
- 专业: 分子生物学
4楼2010-04-16 15:57:28
40