Nuclear와 nucleicacid binding proteins을 분리 정제할 수 있습니다
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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 Kit은/는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료용이 아닙니다.
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Features
- Cytosolic proteins이 없는 nuclear — 원하는 단백질만을 농축 가능함
- Greatly reduced sample complexity
- Highly reproducible fractionation
- Includes protocols for tissue proteomics
Product Details
The Qproteome Nuclear Protein Kit is designed for specific enrichment of nuclear proteins from cultured mammalian cells.
Performance
The Qproteome Nuclear Protein Kit provides highly effective and reproducible separation of nuclear proteins as demonstrated using specific marker proteins (see figure " Reproducible, efficient separation of marker proteins"), which remain functionally active (see figure " Isolation of an active transcription factor").
See figures
Principle
The Qproteome Nuclear Protein Kit is designed for specific enrichment of nuclear proteins from cultured mammalian cells. Lysis and centrifugation are used to separate the cytosolic fraction (supernatant) from the cell nuclei (pellet). A high-salt buffer allows dissociation of nuclear binding proteins (such as transcription factors) and their removal by diffusion from the nuclei.
Procedure
Cells are lysed and centrifuged to isolate nuclei. After washing, an extraction buffer is added to the nuclei and nucleic acid binding proteins dissociate from DNA and RNA. This soluble fraction is separated from the nuclear pellet by centrifugation. Proteins remaining in the pellet are solubilized using a second extraction buffer (see figure " Nuclear protein fractionation procedure").
See figures
Applications
The Qproteome Nuclear Protein Kit delivers a nucleic acid binding protein fraction suitable for a wide range of activity assays.
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.
Specifications
| Features | Specifications |
|---|---|
| ApplicationsKO | 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
Kit Handbooks (1)
Scientific Posters (1)
Safety Data Sheets (2)
Technical Information (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?