For separation of nuclear and nucleic acid binding proteins
Cat. No. / ID: 37582
The Qproteome Nuclear Protein Kit is designed for specific enrichment of nuclear proteins from cultured mammalian cells.
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.
The Qproteome Nuclear Protein Kit delivers a nucleic acid binding protein fraction suitable for a wide range of activity assays.
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.
| 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 |
