Qproteome Nuclear Protein Kit

核蛋白分离和分类抽提

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Qproteome Nuclear Protein Kit

Cat. No. / ID: 37582

For 12 nuclear protein preparations: Buffers, Reagents, Protease Inhibitor Solution, Benzonase

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Qproteome Nuclear Protein Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

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✓ Knowledgeable and professional Product & Technical Support

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Features

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

Product Details

Qproteome Nuclear Protein Kit专为哺乳动物培养细胞的核蛋白制备而设计。

Performance

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""），保持功能活跃（参见""）。

See figures

Reproducible, efficient separation of marker proteins.

Isolation of an active transcription factor.

Principle

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。

Procedure

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""）。

See figures

Nuclear protein fractionation procedure. $Nuclear protein fractionation procedure.$

Applications

Qproteome Nuclear Protein Kit可制备核酸结合蛋白，适合广泛的蛋白活性分析。

Supporting data and figures

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.

$Nuclear protein fractionation procedure.$

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

Resources

For isolation of nuclear and nucleic acid binding proteins from eukaryotic cells and tissues

Download Safety Data Sheets for QIAGEN product components.

For isolation of nuclear and nucleic acid binding proteins from eukaryotic cells and tissues

FAQ

The Qproteome Cell Compartment Kit, the Qproteome Nuclear Protein Kit, the Qproteome Mitochondria Isolation Kit, and the PhosphoProtein Purification Kit are compatible with tissue samples. The respective protocol can be found in the respective handbook.

The Qproteome Mammalian Protein Prep Kit is also compatible with tissue, there is the QIAGEN Supplementary Protocol: Purification of protein from animal tissues using the Qproteome Mammalian Protein Prep Kit and the TissueRuptor™ available. For all these protocols the TissueRuptor is used.

FAQ ID -755

The following protocol is suitable for concentrating and desalting protein samples for downstream applications such as 2D-PAGE:

Add four volumes of ice-cold 100% acetone to the protein fraction and incubate for 15 minutes on ice
Centrifuge for 10 minutes at 12,000 x g in a pre-cooled microcentrifuge at 4°C
Discard the supernatant and air dry the pellet
Resuspend the pellet in an appropriate sample buffer required for the downstream application

You will find this protocol also in the handbooks for QIAGEN's QProteome Protein Fractionation Kits.

FAQ ID -1035