Tandem Ubiquitin Binding Entities (TUBEs) capitalize on the linkage of multiple ubiquitin interacting motifs (UIMs) to generate reagents with high affinity for polyubiquitin. Biotin K63-TUBE binds K63-linked polyubiquitin with a 1000 to 10,000-fold preference for K63 over other linkage types and provides a sensitive and cost-effective tool for determining the abundance of K63-linked polyubiquitin in the cell, or identifying the linkage-type associated with a particular protein of interest. Based on an engineered peptide developed by Sims et al., Biotin K63-TUBE consists of multiple ubiquitin interacting motifs separated by rigid linkers, presumably increasing its affinity to extended K63 polyubiquitin chains. Biotin K63-TUBE is useful for detection and characterization of K63-linked polyubiquitylated proteins by Far Western blotting, affinity purification from lysates and biological fluids, and in situ detection.
Far Westerns with Biotin K63-TUBE 1 require only minor modifications to existing immunoblot protocols, the most notable of which is to avoid subjecting the blot to denaturing conditions, e.g. boiling or autoclaving, which are common treatments used in Western blotting protocols for ubiquitin antibodies. Biotin K63-TUBE substitutes for the anti-ubiquitin IgG as the primary detection reagent. Secondary detection is then accomplished with an avidin or streptavidin enzyme conjugate.
Fluorescein-(UM504F) and TAMRA-(UM504T) labeled versions of K63 TUBE are also available for far Western applications. Please inquire for details.
This product is protected by one or more US or Foreign patents. By purchasing this product, the purchaser agrees to comply with the terms of our Limited Use Label License.
|Formulation||0.5 mg/ml in PBS, pH 7.2|
|Molecular Weight||22,024 Da|
- Detection of K63-linked polyubiquitinylated proteins by ligand (far Western) blotting
- Identification of the polyubiquitin linkage-type of your protein of interest
- Inhibition of K63-dependent processes in lysates
- Purification of K63-linked polyubiquitinylated proteins from cell and tissue lysates using avidin supports
- In situ labeling for detection of K63-linked polyubiquitinylated proteins by histochemistry
- Selective for K63-linked polyubiquitin chains
- Superior to a standard antibody-based detection
- Convenient detection via standardized ligand blotting (far western) techniques
- High sensitivity
- Low cost per blot
- Cost Comparing K63 TUBE to K63-specific Antibodies
- K63 Poster
- Limited use of Label License
 Sims, JJ, Cooper, EM, Scavone, F, Kane, LA, Youle, RJ, Boeke, JD, and Cohen, RE. (2012).“Ubiquitin sensor peptides reveal localization and linkage-type dependence of cellular polyubiquitin signaling.”Nature Methods, 9(3):303-9.
 Hjerpe, R, Aillet, F, Lopitz-Otsoa F, Lang, V, England P, and Rodriguez, MS. (2009). “Efficient protection and isolation of ubiquitylated proteins using tandem ubiquitin-binding entities.” EMBO Rep., 10(11):1250-8.
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