Nuclear/Cytosol Fractionation Kit

Application: Immunoprecipitation (IP)

Catalog no.: ABIN412485

Additional Information

Sample Type: Cell Culture

Components: Cytosol Extraction Buffer A (CEB-A), Cytosol Extraction Buffer B (CEB-B), Nuclear Extraction Buffer (NEB), DTT (1 M), Protease Inhibitor Cocktail (lyophilized)

Description: This Nuclear/Cytosol Extraction Kit provides a complete system that enables the separation of nuclear extract from the cytoplasmic fraction of mammalian cells. The optimized reagents and procedures provided with the kit allow separation of nuclear and cytoplasmic fractions quickly with little or no cross-contaminations. The extracted nuclear and cytoplasmic protein fractions are functional and compatible with downstream assays such as transcriptional activity, RNA splicing, gel shift assay, reporter assays, enzyme activity assays, and Western blotting.

Application Details

Application Notes: Note: Nuclear extract prepared using the above procedure contains proteins in a concentration ~1 mg/ml. If higher concentration is desired, the nuclei can be resuspended in less volume of NEB-Mix (such as 20 μ l) in Step 7. IV. Published References Citing the Product: • DeVries, T.A., et al. (2004) J. Biol. Chem. 279:45603-45612. • Fukuda, N., et al. (2003) J. Biol. Chem. 278:50386-50392.279:45603-45612. • Hasegawa H. et al. (2006) Blood 107: 679-688. • Ishitsuka, K., et al. (2005) Blood 106: 1794-1800. • Kaburagi, Y., et al., (2004) Endocrinology 145: 5862-5874. • Kalin, T. V., et al. (2006) Cancer Res. 66: 1712-1720. • Kanouchi, H., et al. (2005) Protein Sci. 14: 2344-2349. • Kaul, S., et al., (2005) 10.1074/jbc.M500022200. • Kawakami S. et al. (2006) J. Pharmacol. Exp. Ther. 317: 1382-1390. • Kirton J.P. et al. (2006) Circ. Res. 98: 1264-1272. • Kobayashi, S., et al., (2005) Genes Cells 10: 253-260. • Laduron, S., et al. (2004) Nucleic Acids Res. 32:4340-4350. • Li, Q., et al. (2004) J. Biol. Chem. 279:52106-52116. • Loriot, A., et al. (2006) J. Biol. Chem. 10.1074/jbc.M510469200. • Mani, Bernhard., et al. (2006) J. Virol. 80: 1015-1024. • Massaad, C.A., et al. (2004) J. Biol. Chem. 279:54470-54478. • Mori, Y., et al. (2005) J. Virol. 79: 3448-3458. • Ogawa, T., et al. (2005) Cancer Res. 65: 9771-9778. • Ruchalski K. et al. (2006) J. Biol. Chem. 281: 7873-7880. • Takahashi Y. et al. (2006) J. Biol. Chem. 281: 21820- 21826. • Tanito, M., et al. (2005) Invest. Ophthalmol. Vis. Sci. 46: 979-987. • Tsukamoto, H., et al. (2004) J. Biol. Chem. 279:48457-48465. • Unoki M. et al. (2006) J. Biol. Chem. 10.1074/jbc.M606296200. • Wang, Z., et al. (2005) Cancer Res. 65: 8224-8232.

Storage: -20°C

Expiry Date: 12 months

Restrictions: For Research Use only

Publications

Peng, Huang, Wu et al.: "Glutamine protects ischemia-reperfusion induced acute lung injury in isolated rat lungs." in: Pulmonary pharmacology & therapeutics, 2010 (PubMed).

Csepeggi, Jiang, Kojima et al.: "Somatic Cell Plasticity and Niemann-Pick Type C2 Protein: FIBROBLAST ACTIVATION." in: The Journal of biological chemistry, Vol. 286, Issue 3, pp. 2078-87, 2011 (PubMed).

Kommaddi, Dickson, Barker: "Stress-induced expression of the p75 neurotrophin receptor is regulated by O-GlcNAcylation of the Sp1 transcription factor." in: Journal of neurochemistry, 2010 (PubMed).

Maag, Maxwell, Hardesty et al.: "Inositol polyphosphate multikinase is a physiologic PI3-kinase that activates Akt/PKB." in: Proceedings of the National Academy of Sciences of the United States of America, 2011 (PubMed).

Lee, Lee, Kim et al.: "Cross-regulation between oncogenic BRAF(V600E) kinase and the MST1 pathway in papillary thyroid carcinoma." in: PLoS ONE, Vol. 6, Issue 1, pp. e16180, 2011 (PubMed).

Biswas, Joo, Liu et al.: "Endoprotease-Mediated Intracellular Protein Delivery Using Nanocapsules." in: ACS nano, 2011 (PubMed).

Product: Ban, Kozar: "Glutamine protects against apoptosis via downregulation of Sp3 in intestinal epithelial cells." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 299, Issue 6, pp. G1344-53, 2010 (PubMed).

Ishikado, Nishio, Morino et al.: "Low concentration of 4-hydroxy hexenal increases heme oxygenase-1 expression through activation of Nrf2 and antioxidative activity in vascular endothelial cells." in: Biochemical and biophysical research communications, Vol. 402, Issue 1, pp. 99-104, 2010 (PubMed).

Zerin, Kim, Hong et al.: "Quercetin reduces oxidative damage induced by paraquat via modulating expression of antioxidant genes in A549 cells." in: Journal of applied toxicology : JAT, 2012 (PubMed).