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产品简介
| 英文名称 | Nuclear Extraction Kit |
| 中文名称 | 细胞核提取试剂盒 |
| 储存条件 | 短期保存在2-8℃;长期保存在-20℃。 |
| 运输条件 | 冷藏运输 |
| 规格 | 50T 100T |
| 单位 | 盒 |
保存：短期使用可4℃储存，长期保存请置于-20℃。
产品内容：
| 组份 | 50T | 100T |
| Lysis Buffer | 100mL | 200mL |
| Reagent A | 2.5mL | 5mL |
| Medium Buffer | 25mL | 50mL |
| Store Buffer | 5mL | 10mL |
产品简介：
细胞核提取试剂盒用于从动物细胞或组织中分离出完整而纯化的细胞核。适合于动物软组织（肝或脑组织）和硬组织（肌肉）以及培养细胞的细胞核的制备。其制备物产量高，可以被用于细胞凋亡、信号传递、代谢和蛋白组学等的研究。产品不含污染性蛋白酶和核酶，性能稳定。
操作步骤：
1. 样本处理
a. 组织匀浆：称取100~200 mg新鲜组织如肝脏、脑、心肌等，PBS或生理盐水冲洗，洗净血水，滤纸吸干，用剪刀剪为碎块放入小容量玻璃匀浆器内。加入1.0 mL预冷的Lysis Buffer，再加入50μL Reagent A，0℃冰浴中上下研磨组织20次；有未研磨开的组织，可用双层纱布过滤。
b. 培养细胞匀浆：消化细胞，PBS洗涤，800 g 离心5~10 min收集细胞，计数。每次提取需要5 ×107个细胞，加入1.0 mL冰预冷的Lysis Buffer重悬细胞，再加入50μL Reagent A，将细胞悬液转移到小容量玻璃匀浆器内，0℃冰浴研磨细胞20~30次。
2. 将组织或细胞匀浆物转移到1.5mL离心管中，4℃，700g 离心5 min。细胞核沉淀在收集管底部，弃上清。加入0.5 mL冰预冷的Lysis Buffer重悬沉淀。
3. 取另一新离心管内加入0.5 mL Medium Buffer，吸取上一步的重悬液，沿管壁小心地加入到此离心管中，置于Medium Buffer之上，4℃，700 g 离心5min。细胞核沉淀在管底。
4. 弃上清，在细胞核沉淀中加入0.5 mL Lysis Buffer重悬细胞核沉淀，1000g 离心10min ，弃上清，得到较纯的细胞核沉淀。
5. 用50-100μL Store Buffer 或合适的反应缓冲液重悬细胞核沉淀，立即使用或-70℃保存。
注意事项：
1. 为保证获得完整的细胞核，务必做到：第一，全程低温操作；第二，快速；第三，在不破坏亚细胞器的情况下破碎细胞，这是制备细胞核的最关键环节。与组织块相比，培养细胞特别是贴壁培养细胞在用玻璃匀浆器匀浆时较难破壁，因而要选用小容量玻璃匀浆器、间隙严密的研杵上下研磨培养细胞。
2. 以离心力g计算正确的离心速度，不同的离心机可据此精确计算离心速度。
3. 进行Western Blot和2D-胶电泳，可直接加入上样缓冲液裂解细胞核。
转速与离心力换算: G=1.11×10-5×R×(rpm)2
G为离心力，一般以ｇ（重力加速度）的倍数来表示;[rpm]2即：转速的平方;R为半径，单位为厘米。
备注：
以上数据均来自公开文献, Solarbio暂未进行独立验证, 仅供参考。
These protocols are for reference only. Solarbio does not independently validate these
methods.
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