TOLLIP [GST-tagged]

Ubiquigent

Catalog No: UBI-66-1016-050

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Species Reactivity: human  
Source: E. coli  
Formula: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol  
Concentration: 0.5 mg/ml  
Molecular Weight: ~57kDa  
Protein Sequence: Accession number: NP_061882.2. For full protein sequence information download the Certificate of Analysis pdf.  
Quality Control Protein Identification: Confirmed by mass spectrometry.  
Quality Control Activity: Ubiquitin Binding Domain Activity: The ubiquitin chain binding activity of GST-TOM1 and GST-TOLLIP were validated through their ability to capture poly ubiquitylated IRAK1 from a lysate preparation derived from IL-1 stimulated HEK293 cells. GST-TOM1 and GST-TOLLIP were pre-incubated with Glutathione Sepharose 4B for 20 minutes at 4 °C followed by incubation for 2 hours at 4 °C with 2mg IL-1 stimulated HEK293 cell lysate. The binding reaction was then centrifuged and the pellet analysed by SDS-PAGE/ Western blotting (Lanes 1 and 2). These samples were compared alongside GST-TOM1 and GST-TOLLIP binding reactions performed with lysates derived from non-stimulated HEK293 cells (Lanes 3 and 4). Ubiquitylated IRAK1 was identified by Western Blotting using an anti-IRAK1 antibody and such species were observed only in the pellet sample derived from a binding reaction containing wild-type GST-TOM1 or GST-TOLLIP and IL-1 stimulated HEK293 cell lysate (Lanes 1 and 2 respectively).