Qproteome Nuclear Protein Kit
For separation of nuclear and nucleic acid binding proteins
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Qproteome Nuclear Protein Kit
Cat no. / ID. 37582
For 12 nuclear protein preparations: Buffers, Reagents, Protease Inhibitor Solution, Benzonase
This product contains substances regulated under REACH (EC 1907/2006 Annex XIV). The use of this product in the EU is permitted subject to an exemption (Article 56(3)). Please refer to the REACH notification and the SDS of this product, both of which can be found in the “Resources” section of this page, for more information.
The Qproteome Nuclear Protein Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Features
- Nuclear preparations free of cytosolic proteins
- 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 |
|---|---|
| applications | SDS-PAGE, mass spectrometry |
| fractionsisolated | Three fractions |
| species | Mammal |
| samplesize | 5 x 10e6 cells |
| forglycoproteinswhichtypeofglycoproteins | n.d |
| bindingcapacityyield | 200–300 µg |
| startmaterial | Cell cultures |
Resources
Kit Handbooks (1)
Scientific Posters (1)
Safety Data Sheets (1)
Technical Information (1)
Certificates of Analysis (1)