>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Ubiquitin chains vary in length, linkage, and function. K48-linked Non-hydrolyzable Tri-Ubiquitin Chains (Ub3) may be useful for investigating Ubiquitin-binding proteins and exploring the role of unanchored Ubiquitin chains in signaling pathways. Reaction conditions will need to be optimized for each specific application. IMPORTANT: Heating this product in SDS-PAGE buffer or terminating reactions containing this product with heated SDS-PAGE buffer could lead to unexpected, high apparent molecular weight banding or smearing on gels that is not representative of product purity. For optimal results, we recommend incubation in SDS-PAGE buffer + DTT at <40 °C for 20 minutes prior to gel electrophoresis.
E. coli-derived human Tri-Ubiquitin protein
Each Ubiquitin contains a Pro substitution at position 73.
Formulation Lyophilized from a solution in deionized water.
Reconstitution Reconstitute at 2 mg/ml in an aqueous buffer.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
6 months from date of receipt, -20 to -70 °C as supplied.
3 months, -20 to -70 °C under sterile conditions after reconstitution.
With a predicted molecular weight of 26 kDa, tri-Ubiquitin chains are composed of three Ubiquitin monomers that are covalently linked through isopeptide bonds, which typically form between a lysine residue of one Ubiquitin molecule and the C-terminal glycine residue of another Ubiquitin (1). Each human Ubiquitin monomer is 76 amino acids (aa) in length and shares 96% and 100% aa sequence identity with yeast and mouse Ubiquitin, respectively (2). Seven of the 76 aa in Ubiquitin are lysine residues that can participate in poly-Ubiquitin chain formation. Linkage through specific lysine residues is thought to serve as a signal that affects protein degradation, signaling, trafficking, and other cellular processes (3-8).
Linkage specific, non-hydrolyzable tri-Ubiquitin is resistant to the activity of deubiquitinating enzymes (DUB's) that cleave the isopeptide linkage between adjacent Ubiquitin molecules. It can be used to investigate binding interactions between tri-Ubiquitin and proteins that contain elements such as Ubiquitin-associated domains (UBAs) or Ubiquitin-interacting motifs (UIMs). This product may also be useful in exploring the role of unanchored poly-Ubiquitin chains in some signaling pathways.
Scheffner, M. et al. (1995) Nature 373:81.
Sharp, P.M. & W.-H. Li (1987) Trends Ecol. Evol. 2:328.
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