WWP1 [untagged]

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Species Reactivity: Human  

Source: E. coli expression  

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

Concentration: 0.5 mg/ml  

Molecular Weight: ~105 kDa  

Protein Sequence: Accession number: NP_008944. For full protein sequence information download the Certificate of Analysis pdf.  

Quality Control Protein Identification: Confirmed by mass spectrometry.  

Quality Control Activity: E3 ligase assay:  The ubiquitin conjugating activity of WWP1 was validated through its ability to catalyse the generation of polyubiquitin chains in the presence of the E1 activating enzyme His-UBE1, the E2 conjugating enzyme His-UBE2L3 (UbcH7) (several E2s were tested, data generated with this E2 is provided by way of example) and ubiquitin. Incubation of WWP1 for 60 minutes at 37 °C in the presence of ubiquitin, His-UBE1, His-UBE2L3 and ATP (Lane 1) was compared alongside two control reactions with either ATP (Lane 2) or WWP1 (Lane 3) excluded from the reaction. Ubiquitin conjugates were identified by Western blotting using an anti-ubiquitin conjugate antibody and these were observed only in the presence of both ATP and WWP1 (with the exception of one species of approximately 100 MW observable in lane 3).