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描述 概述 A rapid method for the isolation of biologically active proteasomes using affinity matrix beads comprised of a GST-fusion protein containing an ubiquitin-like domain (UbL) bound to GST-agarose. The proteasome subunit proteins can be identified by loading the beads directly onto an SDS-PAGE gel and immunoblotting with subunit specific antibodies. Alternatively, proteasome bound to beads can be used in proteolytic assays using proteasome substrates.
• Utilize Ubiquitin-like domain (Ubl) bound to GST-agarose
• Rapidly isolate biologically active proteasomes
• Affinity mature beads can be directly loaded onto SDS-PAGE gels for proteasome subunit identification
• Proteasome-bound beads are compatible with proteolytic assays using proteasome substrates
产品目录编号 539176 品牌系列 Calbiochem®
应用数据 
Total protein extracts were applied to control and proteasome-binding beads, and incubated for 4 h at 4°C. The matrix was washed three times with Buffer A, suspended in SDS sample buffer, separated in a 10% SDS-polyacrylamide gel, and electroblotted to nitrocellulose. The filter was incubated with antibodies against Rpt1, a subunit in the 19S regulatory particle of the proteasome. The first lane shows the recovery of the proteasome subunit Rpt1 with the proteasome-binding GST-UbL beads. The second lane is a mock-reaction (using GST), while the third lane contains 5% of the input lysate. Similar samples were incubated in reaction buffer to measure proteasome-specific proteolytic activity. The hydrolysis of Suc-Leu-Leu-Val-Tyr-AMC was readily detected in the GST-UbL beads, while the no activity was observed with the GST beads.参考 参考 Chen, L., and Madura, K. 2002. Mol. Cell Biol. 22, 4902.
Chen, L., et al. 2001. EMBO Rep. 2, 933.
Schauber, C., et al. 1998. Nature 391, 715.产品信息 声明 Sold under license of U.S. Patent 6,294,363. 格式 1.0 ml bead suspension 试剂盒包含 UbL Beads Sufficient to process 12.5 to 25 mg of Lysate, Control Glutathione-Agarose Beads, Control Lysate, and a user protocol. 生物信息 化验时间 3-5 h 样品类型 Cell extracts 品种 canine, human, mouse 储存和货运信息 货运代码 Blue Ice Only
毒性 Standard Handling
储存 +2°C to +8°C
存储条件 Upon arrival store entire kit contents at 4°C. 无需冷冻 Yes
补充信息 试剂盒包含 UbL Beads Sufficient to process 12.5 to 25 mg of Lysate, Control Glutathione-Agarose Beads, Control Lysate, and a user protocol.
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