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特点
- 制备不含胞浆蛋白的核蛋白
- 极大降低了样本复杂性
- 高度重复性的核蛋白分离制备
- 同样适用于组织样本
产品详情
Qproteome Nuclear Protein Kit专为哺乳动物培养细胞的核蛋白制备而设计。
绩效
Qproteome Nuclear Protein Kit提供高效和可重复的核蛋白质分离，使用特定的标志蛋白（参见" Reproducible, efficient separation of marker proteins"），保持功能活跃（参见" Isolation of an active transcription factor"）。
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原理
Qproteome Nuclear Protein Kit用于从培养的哺乳细胞中抽提核蛋白。通过裂解和离心，使胞浆（上清）和细胞核（沉淀）分离。高盐缓冲液使得核结合蛋白（如转录因子）溶解，从细胞核上分离。
程序
裂解细胞并离心，以分离细胞核。洗涤沉淀，加入提取液从DNA和RNA中提取核酸结合蛋白。再次离心得到可溶性组分，在沉淀中的蛋白质加入第二种提取液进行进一步提取（参见" Nuclear protein fractionation procedure"）。
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应用
Qproteome Nuclear Protein Kit可制备核酸结合蛋白，适合广泛的蛋白活性分析。
辅助数据和图表
Isolation of an active transcription factor.
A biotinylated DNA oligo containing a specific transcription-factor binding sequence was immobilized on a streptavidin-coated 96-well plate. NX1 extraction buffer (Blank) or 10 µg nucleic acid binding protein fraction was added, washed, and detected colorimetrically in an ELISA procedure using a transcription-factor specific antibody. Specificity of binding was demonstrated by addition of a 10x excess of non-biotinylated oligo that was able to displace the transcription factor.
Specifications
| Features | Specifications |
|---|---|
| Applications | SDS-PAGE, mass spectrometry |
| Fractions isolated | Three fractions |
| Species | Mammal |
| Sample size | 5 x 10e6 cells |
| For glycoproteins: which type of glycoproteins | n.d |
| Binding capacity/yield | 200–300 µg |
| Start material | Cell cultures |
资源
试剂盒操作手册 (1)
学术海报 (1)
安全数据表 (1)
技术资讯 (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
FAQ
Which Qproteome or Protein Fractionation Kits from QIAGEN are compatible with tissue samples?
How do I perform an Acetone Precipitation for concentrating and desalting protein samples?