Millipore
ProteoExtract® Subcellular Proteome Extraction Kit
别名：S-PEK Kit
英文名：
PE Subcellular Proteome Extraction Kit
保存条件：
2-8℃
供应商：
Merck
规格：
20 TESTS
ProteoExtract®亚细胞蛋白质组抽提试剂盒通过2小时的简便操作，高效分离细胞质蛋白、膜蛋白、细胞核蛋白及细胞骨架蛋白等4个亚细胞蛋白质组，特别适合用于蛋白的定位、迁移等分析，为蛋白功能描述和疾病研究提供便利。操作不需要超声、超高速离心等费时费力的处理过程，也不需要昂贵的仪器设备，得率高且保持蛋白天然活性及结构。起始材料仅为10^6个细胞以上；可用于20次抽提（货号为539791的试剂盒用于10次抽提）；下游兼容2D电泳、芯片检测、ELISA、酶活分析及质谱分析等。
另提供配套的WB检测抗体套装，货号71771。
属性
用途
sufficient for 20 extractions
manufacturer/tradename
Calbiochem®
储存条件
OK to freeze
avoid repeated freeze/thaw cycles
input
sample type: mammalian cultured cells
sample type: mammalian tissue(s)
运输
ambient
储存温度
2-8°C
说明
一般描述
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.
包装
1 kit in Glass bottle
特点和优势
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
组分
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.
警告
Toxicity: Multiple Toxicity Values, refer to MSDS (O)
原理
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.
制备说明
Appropriate samples types include:
Kit Components Needed for One Extraction
The volume of each component required for one subcellular extraction depends on the amount of starting material and is shown in the table below.
Table 2: Buffer Volumes Required for S-PEK Extraction Based on Cell Number
*Tested on rat liver and bovine liver tissue.
The following table lists the amount of extraction buffer needed for extracting proteins from cells grown in various size tissue culture flasks and dishes. As guideline to estimate the amount of starting material please refer to the table above for a listing of expected protein yields obtained from selected cell types following S-PEK extraction.<div class="Bio_doc_image">Table 3: Buffer Volumes Required for S-PEK Extraction Based on Flask/Dish Size
储存及稳定性
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.
其他说明
Zhang, L., and Insel, P. A. 2004. J. Biol. Chem.279, 20858.
Yuan, X., et al. 2002. Electrophoresis23, 1185.
Butcher, et al. 2001. J. Immunol.167, 2193.
Ott, et al. 2001. Pharmacogenomics J.1, 142.
Allen, L. 2000. Nature405, 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. Electrophoresis18, 2842.
Laemmli, U. K. 1970. Nature227, 680.
Lowry, et al. 1951. J. Biol. Chem.193, 265.
http://www.expasy.ch/ and http://www.expasy.proteome.org.au
法律信息
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
PROTEOEXTRACT is a registered trademark of Merck KGaA, Darmstadt, Germany
安全信息
储存分类代码
10 - Combustible liquids
