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- 详细信息
- 文献和实验
- 技术资料
- 供应商：
伟进生物
- 库存：
大量
- 英文名：
NE-PER Nuclear and Cytoplasmic Extraction Reagents
- 规格：
kit
Sufficient reagents for extracting 50 cell pellet fractions having packed cell volumes of 20 µl each (a total of approx.
Kit includes:
Cytoplasmic Extraction Reagent I (CER I), 10 ml
Cytoplasmic Extraction Reagent II (CER II), 550 µl
Nuclear Extraction Reagent (NER), 5 ml
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文献和实验Sequential Extraction of Proteins by Chemical Reagents
experimentation. A simple and robust technique for separating highly soluble (hydrophilic) cytoplasmic proteins from poorly soluble (hydrophobic) membrane-associated proteins uses differential solubility in a progressive series of extraction buffers
Preparation of nuclear extract and cytoplasmic extract
Solutions:Buffer A (Hypotonic Buffer): 1L10 mM HEPES pH 7.9 10 ml 1M HEPES,pH 7.91.5 mM MgCl2 1.5 ml 1M MgCl210 mM KCl3.33 ml 3M KCl0.5 mM DTT 500 µl 1M DTT0.2 mM PMSF 1 ml 0.2 M PMSFBuffer B (S100 extraction buffer): 500 ml0.3 M HEPES pH 7.9 150 ml
contains the cytoplasmic fraction. The pellet is the nuclear fraction. 9. Resuspend nuclear pellet in 50 μl complete Cell Extraction Buffer for 30 minutes on ice with vortexing at 10 minute internals. 10. Centrifuge for 30 minutes at 14,000 x g
