UBIQUITIN RESEARCH TOOLS
TUBEs (GST Tagged)
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Tandem Ubiquitin Binding Entities (TUBEs) are high affinity ubiquitin traps that bind to poly-ubiquitin chains with low nanomolar Kd and can be used for capture of total ubiquitylated proteins from cell and tissue lysates. Importantly, TUBEs also protect polyubiquitylated proteins from deubiquitylation and proteasomal degradation during purification. Which TUBE is right for you? GST-TUBE 1 consists of tandem UBA domains from the protein ubiquilin, while GST-TUBE 2 has been derived from the UBAs of human Rad23a. Both proteins are fusions with the affinity tag GST allowing for pulldowns with glutathione resin, as well as containing hexahistidine and V5 epitopes for immunodetection. These proteins display nanomolar affinities towards K48 and K63 tetra-ubiquitin chains. The use of GST-TUBEs allows for the protection of polyubiquitinated chains during cell lysis, maximizing the ability to isolate/enrich for your polyubiquitinated protein of interest. Additional members of the TUBE product line: Agarose-TUBEs provide the convenience of "one-step" pulldowns for identification and characterization of polyubiquitin proteins by Western blotting, as well allowing for the isolation of proteins from TUBEs for downstream proteomic studies by mass spectroscopy. Biotin-TUBEs are a sensitive detection reagent for polyubiquitin by ligand blotting ("Far Western"), providing a superior alternative the traditional anti-ubiquitin antibodies. Fluorescent TUBEs are HIS6-TUBEs conjuagted to either fluorescein or TAMRA for cytochemical staining by fluorescent microscopy. The nature of TUBEs allows for efficient detection of polyubiquitylated proteins in their native state, while the versatility of TUBEs meets a wide range of experimental needs. |
