Di-ubiquitin (K29-linked) [untagged]

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Species Reactivity: human  

Source: synthetic/chemical ligation  

Formula: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol  

Concentration: 0.5 mg/ml  

Molecular Weight: ~17.1 kDa  

Protein Sequence: Accession number: P62987. For full protein sequence information download the Certificate of Analysis pdf.  

Quality Control Protein Identification: The linkage type (K29) was confirmed by tandem mass spectrometry. A small (~10%) amount of K27 linkage was also detected in the sample mass spectrometry data.  

Quality Control Activity: Di-ubiquitin cleavage assay: The capacity of the di-ubiquitin substrate to be cleaved was tested using a promiscuous - with respect to ubiquitin linkage specificity - deubiquitylase (GST-USP2). Incubation of the di-ubiquitin for 1 hour at 37 °C was compared either in the absence (Lane 2) or presence (Lane 3) of GST-USP2. The reaction products were compared alongside two control samples containing either mono-ubiquitin (Lane 4) or GST-USP2 (Lane 5) only. Cleavage of the di-ubiquitin and generation of mono-ubiquitin was determined by running reactions on a 4-12% ­SDS-PAGE gel and staining with InstantBlueâ„¢ (Lane 1; molecular weight markers).