Blue Loading Buffer Pack

• 产品编号：CST-7722S      品牌：CST       原厂货号：7722S
• 产品分类：生化试剂 > 生物缓冲液 > 蛋白生化分析缓冲液 > 蛋白质上样缓冲液（Loading buffer)
• 应用分类：

描述：

蓝色上样缓冲液用于western blot中裂解细胞。1. 加1/10 体积30X DTT 还原剂到 1 体积3X SDS 上样缓冲液配制成新鲜的3X 还原 SDS 上样缓冲液。 2. 用双蒸水将3X 还原 SDS 上样缓冲液稀释至1X溶液。本产品可以稀释24ml 1X溶液。 3. 将培养体系中的培养液吸出，用1X PBS洗涤细胞，吸出。 4. 加入1X 还原 SDS 上样缓冲液裂解细胞（六孔板中每孔加入100ul或者10cm2板中加入500ul），立刻将细胞从板上刮掉，将提取物转移至离心管中，至于冰上。 5. 超声10-15分钟使DNA断裂，减少样本的粘度。 6. 于95-100度加热20ul样本，至于冰上降温 7. 离心2-5分钟 8. 向SDS-PAGE中上样20ul（10cmX10cm）

原厂资料：

Description

The Blue Loading Buffer Pack is used to lyse cells for Western blotting analysis

Directions for Use

1. Prepare fresh 3X Reducing SDS Loading Buffer by adding 1/10 volume 30X DTT Reducing Agent to 1 volume of 3X SDS Loading Buffer. 
 2. Dilute 3X SDS Loading Buffer to a 1X solution using ddH2O. This product supplies enough 3X material to make 24ml of 1X solution.      
 3. Aspirate media from cultures; wash cells with 1X PBS; aspirate. 
 4. Lyse cells by adding 1X SDS Loading Buffer (100 µl per well of 6-well plate or 500 µl per plate of 10 cm2 plate). Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Keep on ice. 
 5. Sonicate for 10-15 seconds to shear DNA and reduce sample viscosity.  
 6. Heat a 20 µl sample to 95-100ºC for 5 minutes; cool on ice. 
 7. Microcentrifuge for 2-5 minutes. 
 8. Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).

注意事项：

Storage: Reagents are stored at different temperatures
(see “Solutions and Reagents”).