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ITCH / AIP4 Antibody, Mouse MAb

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Human ITCH Antibody Product Information
Immunogen:Recombinant Human ITCH / AIP4 protein (Catalog#11131-HNCE)
Clone ID:01
Ig Type:Mouse IgG1
Formulation:0.2 μm filtered solution in PBS with 5% trehalose
Preparation:This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, recombinant Human ITCH / AIP4 (rh ITCH / AIP4; Catalog#11131-HNCE; NP_113671.3; Arg526-Glu903). The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography.
Human ITCH Antibody Usage Guide
Specificity:Human ITCH / AIP4
Application:ICC/IF, IF, FCM

FCM: 0.5-2 μg/Test

ICC/IF: 10-25 μg/mL

Storage:This antibody can be stored at 2℃-8℃ for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20℃ to -80℃. Preservative-Free.
Sodium azide is recommended to avoid contamination (final concentration 0.05%-0.1%). It is toxic to cells and should be disposed of properly. Avoid repeated freeze-thaw cycles.
Human ITCH Antibody IF Application Image
ITCH / AIP4 Antibody, Mouse MAb, Immunofluorescence
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Immunofluorescence staining of Human ITCH in Hela cells. Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with Mouse anti-Human ITCH monoclonal antibody (15 µg/ml) at 37℃ 1 hour. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (green) and counterstained with DAPI (blue). Positive staining was localized to cytoplasm.
Human ITCH Antibody FC Application Image
ITCH / AIP4 Antibody, Mouse MAb
[Click to enlarge image]
Flow cytometric analysis of Human ITCH expression on HeLa cells. The cells were treated according to manufacturer’s manual (BD Pharmingen™ Cat. No. 554714), stained with purified anti-Human ITCH, then a FITC-conjugated second step antibody. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells.
Other ITCH Antibody Products
ITCH/AIP4 Background

E3 ubiquitin-protein ligase Itchy homolog, also known as Atrophin-1-interacting protein 4, NFE2-associated polypeptide 1, NAPP1 and ITCH, is a cell membrane protein which contains one C2 domain, one HECT (E6AP-type E3 ubiquitin-protein ligase) domain and contains four WW domains. ITCH acts as an E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. It catalyzes 'Lys-29'-, 'Lys-48'- and 'Lys-63'-linked ubiquitin conjugation. ITCH is involved in the control of inflammatory signaling pathways. It is an essential component of a ubiquitin-editing protein complex, comprising also TNFAIP3, TAX1BP1 and RNF11, that ensures the transient nature of inflammatory signaling pathways. ITCH promotes the association of the complex after TNF stimulation. Once the complex is formed, TNFAIP3 deubiquitinates 'Lys-63' polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains. This leads to RIPK1 proteosomal degradation and consequently termination of the TNF- or LPS-mediated activation of NFKB1. Defects in ITCH are the cause of syndromic multisystem autoimmune disease (SMAD) which is characterized by organomegaly, failure to thrive, developmental delay, dysmorphic features and autoimmune inflammatory cell infiltration of the lungs, liver and gut.

Human ITCH/AIP4 References
  • Marchese A. et al., 2003, Dev. Cell 5:709-22.
  • Wang Y. et al., 2006, EMBO J. 25: 5058-70.
  • Bhandari D. et al., 2009, Mol. Biol. Cell 20:1324-39.
  • Edwards TL. et al., 2009, Biochem. J. 423:31-9.
  • Zhang P. et al., 2010, J. Biol. Chem. 285:8869-79.
  • Azakir B.A. et al., 2010, FEBS J. 277:1319-30.
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    Catalog: 11131-MM01-50
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