Biolegend, 611451, Purified anti-ITCH Antibody, 25μg

ITCH, Itchy E3 Ubiquitin Protein Ligase or atrophin-1 interacting protein 4 (AIP4), receives ubiquitin from E2 ubiquitin-conjugating enzymes and transfers them to their target proteins for lysosomal degradation. The degradation regulated by ITCH is involved in a broad array of biological functions, including T helper cell inflammatory signaling, T lymphocyte differentiation, immune response, tumor cell death, and skin keratinocyte differentiation. In animal models, ITCH mutant mice develop spontaneous autoimmune disease symptoms including skin scratching, lymphoproliferation, and chronic pulmonary inflammation. ITCH suppresses IL-4 levels by degrading JUNB and maintains the attenuation of differentiation of T helper cells type 2. To maintain peripheral immune tolerance, ITCH monoubiquitinates protein kinase θ and phospholipase C-γ1 through lysosome degradation near immunological synapse for T cell energy. ITCH modulates trafficking of receptor CXCR4 from the cell surface to the endosome through endocytosis. The transcription factor targets of ITCH, such as JUN, JUNB, TP63, NOTCH1, and GLI1, participate in epidermal keratinocyte development and differentiation. In addition to a role in proteolysis, the polyubiquitination of BRAF by ITCH sustains its activity and elevates the MEK/ERK signaling pathway to promote proliferation and invasion in melanoma cells.
25μg
Verified Reactivity: Human, Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: Recombinant fragment of human ITCH
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application: WB - Quality tested ICC - Verified
Recommended Usage: Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.25 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: For immunocytochemistry (ICC), we suggest fixation with 4% PFA followed by permeabilization with ice-cold methanol or fixation and permeabilization with ice-cold methanol. Fixation with 4% PFA followed by permeabilization with Triton X-100 results in dimmer staining in comparison to the other two methods. Clone W19229A is not suitable for IHC.
RRID: AB_2936758 (BioLegend Cat. No. 611451) AB_2936758 (BioLegend Cat. No. 611452)
Structure: ITCH is a 903 amino-acid protein with a predicted molecular weight of 102.8 kD
Distribution: Ubiquitous; localized to the membrane, cytoplasm, nucleus, trans-Golgi network, endosome, and lysosome
Function: E3 ubiquitin-protein ligase transferring ubiquitin to target protein
Interaction: JUN, JUNB, CXCR4, FYN, ERRB4, protein kinase θ, phospholipase C-γ1 and virus proteins
Cell Type: B cells, Basophils, Dendritic cells, Eosinophils, Lymphocytes, Neutrophils, NK cells, T cells, Th2, Tregs
Biology Area: Adaptive Immunity, Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Death, Immunology, Innate Immunity, Protein Trafficking and Clearance, Signal Transduction, Stem Cells, Ubiquitin/Protein Degradation
Molecular Family: Endosomal Markers, Enzymes and Regulators, Innate Immune Signaling, Ligases, Lysosomal Markers
Antigen References: Marchese A, et al. 2003. Dev Cell. 5:709-22. Chastagner P, et al. 2008. PLoS One. 3:e2735 Sundvall M, et al. 2008. Proc Natl Acad Sci U S A. 105:4162-7. Yin Q, et al. 2019. Nat Commun. 10:1870.
Gene ID: 83737
UniProt: View information about ITCH on UniProt.org
Clone: W19229A
Regulatory Status: RUO
Other Names: ITCHY, Atrophin-1-interacting protein 4, AIP4, HECT-type E3 ubiquitin transferase Itchy homolog, NFE2-Associated Polypeptide 1, NAPP1, AIF4, ADMFD
Isotype: Rat IgG2b, κ