| Description |
The mammalian Itchy homolog, or ITCH, (also known as Atrophin-1-interacting protein 4 or AIP4) is a HECT domain class ubiquitin E3 ligase. ITCH ubiquitinylates the phosphorylated form of Dishevelled protein and promotes its degradation via the Ubiquitin Proteasome System, thereby inhibiting canonical Wnt signaling. The absence of ITCH has been shown to cause severe autoimmune disease in mice. Recent studies have identified multisystem autoimmune disease and morphologic and developmental abnormalities in human patients with ITCH deficiency, thus underscoring the importance of ITCH in many cellular processes. This ligase may catalyze K29-, K48-, and/or K63-linked polyubiquitin chain formation on a variety of reported targets. This kit is designed for in vitro ITCH-mediated ubiquitinylation of user-supplied substrates. Ubiquitinylated proteins can be used in downstream applications, or analyzed by Western blot using antibodies specific for the target protein. A detection antibody is included in the kit to monitor autoubiquitinylation of ITCH. Kit contains reagents sufficient for 10 x 30 μl reactions and 5 Western Blots (mini-gel format).
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