IPAH9.8 [GST-tagged]

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Species Reactivity: Shigella flexneri  

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: ~88.8 kDa  

Protein Sequence: Accession number: AAP79029.1. 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 GST-IpaH9.8 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-UBE2D2 (UbcH5b) (several E2s were tested, data generated with this E2 is provided by way of example) and ubiquitin. Incubation of GST-IpaH9.8 for 30 minutes at 30 °C in the presence of ubiquitin, His-UBE1, His-UBE2D2 and ATP (Lane 1) was compared alongside two control reactions with either ATP (Lane 2) or GST-IpaH9.8 (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 GST-IpaH9.8.