用途
sufficient for 20 extractions
制造商/商品名称
Calbiochem®
储存条件
OK to freeze
avoid repeated freeze/thaw cycles
技术
fractionation: suitable
输入
sample type: mammalian tissue(s)
sample type: mammalian cultured cells
运输
ambient
储存温度
2-8°C
一般描述
哺乳动物细胞亚细胞蛋白质组的快速、可重复提取
ProteoExtract®亚细胞蛋白质组提取试剂盒(S-PEK)设计用于从贴壁和悬浮生长的哺乳动物细胞中,快速且可重复地提取亚细胞蛋白质组。S-PEK利用了四种选定试剂中某些亚细胞室的不同溶解度。对于贴壁细胞,该程序直接在组织培养皿中进行,无需去除细胞。在亚细胞室的连续提取过程中,细胞或细胞的部分仍然附着在平板上,直到使用适当的提取试剂。因此,可以防止细胞从组织培养板上酶促或机械分离,以及不同亚细胞室的任何混合而导致细胞结构的早期破坏。对于悬浮培养的细胞,提取从温和沉淀和细胞清洗开始。分步提取可从一个样品中提取四种不同的蛋白质组分:
样品大小:3-5x106或25-50 mg组织。
ProteoExtract®亚细胞蛋白质组提取试剂盒(S-PEK)设计用于从贴壁和悬浮生长的哺乳动物细胞中,快速且可重复地提取亚细胞蛋白质组。S-PEK利用了四种选定试剂中某些亚细胞室的不同溶解度。对于贴壁细胞,该程序直接在组织培养皿中进行,无需去除细胞。在亚细胞室的连续提取过程中,细胞或细胞的部分仍然附着在平板上,直到使用适当的提取试剂。因此,可以防止细胞从组织培养板上酶促或机械分离,以及不同亚细胞室的任何混合而导致细胞结构的早期破坏。对于悬浮培养的细胞,提取从温和沉淀和细胞清洗开始。分步提取可从一个样品中提取四种不同的蛋白质组分:
- 细胞质部分(F1)
- 膜/细胞器蛋白质部分(F2)
- 核蛋白部分(F3)
- 细胞骨架部分(F4)
样品大小:3-5x106或25-50 mg组织。
特点和优势
- 从一个样品中逐步提取出四种不同的亚细胞蛋白质组。
- 高重复性
- 的无超速离心步骤
- 快速—仅需2小时,手动操作时间为 45 分钟,
- 即可生产出适合功能研究的蛋白质
组分
洗涤缓冲液、提取缓冲液I、提取缓冲液II、提取缓冲液III、提取缓冲液IV、蛋白酶抑制剂混合物、苯甲酸酶®核酸酶(目录号71206)和用户协议。
警告
毒性:多个毒性值,请参阅MSDS(O)
原理
Calbiochem ProteoExtract亚细胞蛋白质组提取试剂盒用于从贴壁组织培养细胞、悬浮组织培养细胞、冷冻细胞颗粒和碎片组织的胞质、细胞器和膜、细胞核和细胞骨架部分亚细胞提取哺乳动物蛋白质。
制备说明
一次提取所需的试剂盒组件
一次亚细胞提取所需的每种组分的体积取决于起始原料量。
一次亚细胞提取所需的每种组分的体积取决于起始原料量。
合适的样品类型包括:
- 贴壁组织培养细胞
- 混悬培养的组织培养细胞
- 冷冻细胞颗粒
- 碎片组织
储存及稳定性
在执行提取方案之前,所有冷冻缓冲液必须在室温下解冻。将水浴温度设置为25°C将有助于解冻过程。解冻后,轻轻摇动或旋转,充分混合缓冲液。
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
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
危险分类
Aquatic Chronic 3 - Eye Irrit. 2 - Skin Irrit. 2
WGK
WGK 3
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