TUBEs: A Revolution inPolyubiquitin Isolation and Characterization
The use ofTandem Ubiquitin Binding Entities (TUBEs) is emerging as an indispensablestrategy for ubiquitin research (1, 2). The first generation of these TUBEsbind K48- and K63-linked tetraUb chains with single digit nanomolar Kds, ~100to 1000-fold more tightly than monomeric UBAs. TUBEs also protect proteins fromDUBs and the proteasome, even in the absence of inhibitors normally required toblock such activity. This allows efficient isolation of native polyUb chainsand attached proteins from cell lines, tissues, and organs under conditionsthat are less likely to alter cell physiology than those listed above. TUBE1and TUBE2 have recently been demonstrated to enrich for all polyUb chainlinkage types, without discrimination, making these reagents appropriate evenif the linkage type is not known for the protein of interest (1).
The next generation of TUBEs: Linkage Specific Isolation of PolyUb Chains
K63 TUBEwas developed to show enhanced selectivity for K63-linked polyubiquitin chains(~20 nM) over all other linkages (>2 µM). It can be used alone or inconjunction with our other TUBE products, especially K48 TUBE HF and M1(linear) TUBE to investigate polyubiquitin chain linkage in your substrateprotein.
Magnetic-TUBEs are TUBEmoieties directly coupled to magnetic beads, for the identification andcharacterization of polyubiquitinated proteins by western blotting and/ordownstream proteomic studies. Magnetic-TUBEs facilitate convenient"one-step" pull-down of polyubiquitinated proteins.
Info
K63 TUBE, Magnetic
Size:1ml
Buffer:PBS, pH 7.2
Storage:Stable for up to 6 months at 4oC.Do not centrifuge at high speeds (>3000g), dry orfreeze the beads.
APPLICATIONS
· Isolation and enrichment of K63-polyubiquitinated proteins from cell andtissue extracts
· Isolate K63-polyubiquitinated proteins for proteomic studies
BENEFITS
· 100-fold preference for K63 polyubiquitin chains over all other linkages
· TUBEs display higher affinity towards polyubiquitin than most ubiquitinantibodies
· Avoids overexpression of epitope-tagged ubiquitin for pull downs
Documents
REFERENCES
- Hjerpe, R., Aillet, F., Lopitz-Otsoa, F., Lang, V., England, P., and Rodriquez, M. S. (2009) Efficient protection and isolation of ubiquitylated proteins using tandem ubiquitin-binding entities, EMBO Rep 10, 1250-1258.
- Hjerpe, R., and Rodriguez, M. S. (2008) Efficient approaches for characterizing ubiquitinated proteins, Biochem Soc Trans 36, 823-827.
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那糖酵解中依赖NAD的酶不是要在线粒体内反应？
蛋白酶分布广泛,主要存在于人和动物消化道中，而消化道通过口与肛门与外界相通，属于外环境或外界环境
（溶菌酶、凝血酶原等）分布在内环境
（消化）酶分布在消化道
