merck millipore,默克密理博,539790,ProteoExtract Subcellular Proteome Extraction Kit

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merck millipore,默克密理博,539790,ProteoExtract Subcellular Proteome Extraction Kit
产品名称：ProteoExtract Subcellular Proteome Extraction Kit
产品型号：539790

merck millipore,默克密理博,539790,ProteoExtract Subcellular Proteome Extraction Kit

产品介绍

merck millipore,默克密理博,539790,ProteoExtract Subcellular Proteome Extraction Kit

描述
概述	Fast and reproducible extraction of subcellular proteomes from mammalian cells ProteoExtract^® Subcellular Proteome Extraction Kit (S-PEK) is designed for fast and reproducible extraction of subcellular proteomes from adherent and suspension-grown mammalian cells. The S-PEK takes advantage of the different solubilities of certain subcellular compartments in the four selected reagents. In the case of adherent cells, the procedure is performed directly in the tissue culture dish without the need for cell removal. Cells or the parts of the cells remain attached to the plate during sequential extraction of subcellular compartments, until the appropriate extraction reagent is used. Thus, the early destruction of the cellular structure by enzymatic or mechanical detachment of cells from the tissue culture plate and any mixing of different subcellular compartments is prevented. For suspension-grown cells, extraction starts with gentle sedimentation and washing of the cells. The stepwise extraction delivers four distinct protein fractions from one sample: Cytosolic fraction (F1) Membrane/organelle protein fraction (F2) Nucleic protein fraction (F3) Cytoskeletal fraction (F4) Proteins are obtained in the native state making the S-PEK suitable for many downstream applications such as 1D and 2D gel electrophoresis, immunoblotting, enzyme activity assays, and protein microarrays. Sample size: 3-5x10⁶ or 25-50 mg tissue.
产品目录编号	539790
品牌系列	Calbiochem®
同义词	S-PEK Kit
特点及优点	Stepwise extraction resulting in four distinct subcellular proteomes from one sample Highly reproducible No ultracentrifugation steps Fast—needs just 2 hours with 45 minutes hands-on time Produces proteins suitable for functional studies
应用数据	A: Adherent SAOS cells were extracted according to the Detailed Protocol for Subcellular Extraction of Proteins From Adherent Cells as outlined above. Images i-iv show the morphology of the cells before and after each extraction step (200-fold enlarged). The SAOS cells remained attached throughout the extraction procedure. B: An aliquot of each fraction from A were subjected to SDS-PAGE analysis (F1-F4 = fractions 1-4). The data demonstrate clear differences in the protein banding patterns among the 4 fractions. C: Aliquots of each fraction from A were separated by SDS-PAGE and transferred to PVD membrane for blotting with the indicated antibodies. For c-Fos, the fractions were subjected to immunoprecipitation, prior to Western blotting, to enrich for any c-Fos present in each fraction. The data demonstrate that each marker protein is specifically enriched within the appropriate fraction. A431 cells were stimulated with 0.2 µg/ml TNF-α for the indicated times. At the end of each induction period the cells were extracted as outlined in the Detailed Protocol for Subcellular Extraction of Proteins from Adherent Cells. The proteins from an aliquot of each fraction were separated by SDS-PAGE and transferred to PVD membrane for Western blot analysis using an antibody specific for NF-κB. The data indicate that there is measurable translocation of NF-κB from the cytoplasm to the nucleas as early as 5 min after TNF-α stimulation. *Tested on rat liver and bovine liver tissue.

参考
参考	Zhang, L., and Insel, P. A. 2004. J. Biol. Chem. 279, 20858. Yuan, X., et al. 2002. Electrophoresis 23, 1185. Butcher, et al. 2001. J. Immunol. 167, 2193. Ott, et al. 2001. Pharmacogenomics J. 1, 142. Allen, L. 2000. Nature 405, 819. Dunn, M. J. 2000. Electrophoresis 6. Rabilloud, T. 2000. Two-dimensional Gel Electrophoresis and Identification Methods Springer-Verlag Mejdoubi, et al. 1999. Biochem. Biophys. Res. Comm. 254, 93. Reymond, et al. 1997. Electrophoresis 18, 2842. Laemmli, U. K. 1970. Nature 227, 680. Lowry, et al. 1951. J. Biol. Chem. 193, 265.

参考

Zhang, L., and Insel, P. A. 2004. J. Biol. Chem. 279, 20858.
Yuan, X., et al. 2002. Electrophoresis 23, 1185.
Butcher, et al. 2001. J. Immunol. 167, 2193.
Ott, et al. 2001. Pharmacogenomics J. 1, 142.
Allen, L. 2000. Nature 405, 819.
Dunn, M. J. 2000. Electrophoresis 6.
Rabilloud, T. 2000. Two-dimensional Gel Electrophoresis and Identification Methods Springer-Verlag
Mejdoubi, et al. 1999. Biochem. Biophys. Res. Comm. 254, 93.
Reymond, et al. 1997. Electrophoresis 18, 2842.
Laemmli, U. K. 1970. Nature 227, 680.
Lowry, et al. 1951. J. Biol. Chem. 193, 265.

产品信息
形式	20 Extractions
试剂盒包含	Wash buffer, Extraction Buffer I, Extraction Buffer II, Extraction Buffer III, Extraction Buffer IV, Protease Inhibitor Cocktail, Benzonase^® Nuclease (Cat. No. 71206), and a user protocol.

应用
应用参考	Sabio, G., et al. 2005. EMBO J. in press. Efanov, A.M., et al. 2004. Diabetes 53, s75. Singh, L.P., et al. 2004. Am. J. Physiol. Renal Physiol. 286, F409.

生物信息
样品类型	Mammalian tissue, cultured mammalian cells

安全信息
R相	R: 22-36/38-52/53 Harmful if swallowed. Irritating to eyes and skin. Harmful to aquatic organisms, may cause long-term adverse effects in the aquatic environment.
S相	S: 22-26-28.2-36/37-45 Do not breathe dust. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. After contact with skin, wash immediately with soap and plenty of water. Wear suitable protective clothing and gloves. In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).

产品使用声明
预定用途	The Calbiochem^® ProteoExtract^® Subcellular Proteome Extraction Kit is designed for the subcellular extraction of mammalian proteins from the cytosolic, organelle and membrane, nuclear, and cytoskeletal fractions of adherent tissue culture cells, suspension tissue culture cells, frozen cell pellets, and fragmented tissues.

储存和货运信息
货运代码	Ambient Temperature Only
毒性	Multiple Toxicity Values, refer to MSDS
储存	+2°C to +8°C
存储条件	Upon arrival store the components of the kit under the following conditions: Table 1: Storage Information *Prior to performing the extraction protocol all frozen buffers must thawed at room temperature. A water bath set at 25°C will aid in the thawing process. After thawing, mix the buffers well gentle shaking or vortexing.
避免冻结 / 解冻	Avoid freeze/thaw
无需冷冻	No

补充信息
试剂盒包含	Wash buffer, Extraction Buffer I, Extraction Buffer II, Extraction Buffer III, Extraction Buffer IV, Protease Inhibitor Cocktail, Benzonase^® Nuclease (Cat. No. 71206), and a user protocol.