Rabbit ITCH Polyclonal Antibody | anti-ITCH antibody

Phospho-ITCH (Tyr420) Antibody

Cat. Num. AAA9613128

• Gene Names: ITCH; AIF4; AIP4; NAPP1; dJ468O1.1
• Reactivity: Human, Mouse; Predicted Reactivity: Pig (100%), Bovine (100%), Horse (100%), Rabbit (100%), Dog (100%), Chicken (91%)
• Applications: Western Blot, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: ITCH; Polyclonal Antibody; Phospho-ITCH (Tyr420) Antibody; ADMFD; AIF4; AIP4; Atrophin 1 interacting protein 4; Atrophin-1-interacting protein 4; dJ468O1.1; dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4)); dJ468O1.1 atrophin 1 interacting protein 4 AIP4; E3 ubiquitin protein ligase Itchy homolog; E3 ubiquitin-protein ligase Itchy homolog; EC 6.3.2; Itch; ITCH_HUMAN; Itchy E3 ubiquitin protein ligase; Itchy E3 ubiquitin protein ligase homolog; Itchy E3 ubiquitin protein ligase homolog mouse; mouse; homolog of; Itchy homolog E3 ubiquitin protein ligase; Itchy mouse homolog E3 ubiquitin protein ligase; NAPP1; NFE2 associated polypeptide 1; NFE2-associated polypeptide 1; Ubiquitin protein ligase ITCH; anti-ITCH antibody

• Host: Rabbit
• Reactivity: Human, Mouse; Predicted Reactivity: Pig (100%), Bovine (100%), Horse (100%), Rabbit (100%), Dog (100%), Chicken (91%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-ITCH (Tyr420) Antibody detects endogenous levels of ITCH only when phosphorylated at Tyr420.; Tissue Specificity: Widely expressed.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-ITCH antibody: Western Blot (WB), Peptide ELISA (EIA)
• Application Notes: WB: 1:500-1:2000; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human ITCH around the phosphorylation site of Tyr420.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: On T-cell activation, phosphorylation by the JNK cascade on serine and threonine residues surrounding the PRR domain accelerates the ubiquitination and degradation of JUN and JUNB. The increased ITCH catalytic activity due to phosphorylation by JNK1 may occur due to a conformational change disrupting the interaction between the PRR/WW motifs domain and the HECT domain and, thus exposing the HECT domain (By similarity). Phosphorylation by FYN reduces interaction with JUNB and negatively controls JUN ubiquitination and degradation.Monoubiquitinated. Autopolyubiquitinated with 'Lys-63' linkages which does not lead to protein degradation.
• Subunit Structure: Monomer. Interacts (via WW domains) with OCNL (By similarity). Interacts (via WW domains) with NOTCH1 (By similarity). Interacts (via WW domains) with JUN (By similarity). Interacts with JUNB; the interaction promotes ITCH-mediated ubiquitination and degradation of JUNB. Interacts with FYN; the interaction phosphorylates ITCH on Tyr-420 decreasing binding of JUNB. Interacts (via WW domain 2) with N4BP1; the interaction inhibits the E3 ubiquitin-protein ligase activity (By similarity). Interacts with NDFIP1 and NDFIP2; this interaction activates the E3 ubiquitin-protein ligase and may induce its recruitment to exosomes (By similarity). Interacts with ARHGEF7. Interacts with RNF11. Interacts (via the WW 1 domain) with NFE2 (via the PXY motif 1); the interaction promotes 'Lys-63'-linked ubiquitination of NFE2, retains it in the cytoplasm and prevents its transactivation activity. Interacts (via WW domains) with CXCR4 (via C-terminus); the interaction depends on CXCR4 phosphorylation. Found in a complex with E3 ligase DTX3L and ESCRT-0 components HGS and STAM. Interacts with DTX3L (via C-terminus); the interaction is increased upon CXCL12 stimulation and inhibits ITCH catalytic activity (the interaction is direct). Interacts with HGS. Interacts (via WW domains) with PCBP2 within a complex containing ITCH, MAVS and PCBP2. Interacts (via WW domains) with TXNIP (via C-terminus). Interacts with p15 BID. Interacts with ERBB4. Interacts with DTX1. Interacts with SPART. Interacts with SNX9 and SNX18. Interacts (via its WW domains) with ATN1. Interacts (via WW domains) with SGK3. Interacts with CBLC. Interacts with OTUD7B. Interacts (via WW domain 1,2 and 3) with PI4K2A; the interaction inhibits PI4K2A catalytic activity and promotes ITCH catalytic activity. Interacts with ARRDC4. Part of a complex containing ITCH, NDFIP1 and MAP3K7 (By similarity). Interacts with UBE2L3; the interaction is mediated by NDFIP1. Interacts with MAPK8/JNK1 (By similarity). Interacts (via WW domains) with ARRDC1 (via PPxY motifs); the interaction is direct and participates to the recruitment of the ubiquitin-protein ligase ITCH to the NOTCH1 receptor. Interacts (via WW domains) with ARRDC2. Interacts (via WW domains) with ARRDC3. (Microbial infection) Interacts with Epstein-Barr virus LMP2A. (Microbial infection) Interacts with Human cytomegalovirus (HCMV) protein UL42; this interaction induces ubiquitination and degradation of ITCH. (Microbial infection) Interacts with herpesvirus 1 (HHV-1) UL56 protein; this interaction induces ubiquitination and probably degradation of ITCH. (Microbial infection) Interacts with herpesvirus 2 (HHV-2) UL56 protein. (Microbial infection) Interacts with varicella-zoster virus (VZV) Orf0 protein. (Microbial infection) Interacts with herpesvirus 6A (HHV-6A) U24 protein. (Microbial infection) Interacts with ebola virus protein VP40; this interaction is required for efficient viral egress from the infected cell.
• Similarity: The WW domains mediate interaction with PPxY motif-containing proteins.
• Subcellular Location: Cell membrane>Peripheral membrane protein>Cytoplasmic side. Cytoplasm. Nucleus. Early endosome membrane>Peripheral membrane protein>Cytoplasmic side. Endosome membrane>Peripheral membrane protein>Cytoplasmic side.; Note: May be recruited to exosomes by NDFIP1 (PubMed:18819914). Localizes to plasma membrane upon CXCL12 stimulation where it co-localizes with CXCL4 (PubMed:14602072). Localization to early endosomes is increased upon CXCL12 stimulation where it co-localizes with DTX3L and CXCL4 (PubMed:24790097).
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis of extracts from NIH 3T3, using Phospho-ITCH(Tyr420) Antibody. Lane1 was treated with phospho-blocking peptide, Lane2 was treated with non-phospho-blocking peptide.)

Related Product Information for anti-ITCH antibody

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. Catalyzes 'Lys-29'-, 'Lys-48'- and 'Lys-63'-linked ubiquitin conjugation. Involved in the control of inflammatory signaling pathways. Essential component of a ubiquitin-editing protein complex, comprising also TNFAIP3, TAX1BP1 and RNF11, that ensures the transient nature of inflammatory signaling pathways. 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 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NFKB1. Ubiquitinates RIPK2 by 'Lys-63'-linked conjugation and influences NOD2-dependent signal transduction pathways. Regulates the transcriptional activity of several transcription factors, and probably plays an important role in the regulation of immune response. Ubiquitinates NFE2 by 'Lys-63' linkages and is implicated in the control of the development of hematopoietic lineages. Mediates JUN ubiquitination and degradation (By similarity). Mediates JUNB ubiquitination and degradation. Critical regulator of type 2 helper T (Th2) cell cytokine production by inducing JUNB ubiquitination and degradation (By similarity). Involved in the negative regulation of MAVS-dependent cellular antiviral responses. Ubiquitinates MAVS through 'Lys-48'-linked conjugation resulting in MAVS proteasomal degradation. Following ligand stimulation, regulates sorting of Wnt receptor FZD4 to the degradative endocytic pathway probably by modulating PI42KA activity. Ubiquitinates PI4K2A and negatively regulates its catalytic activity. Ubiquitinates chemokine receptor CXCR4 and regulates sorting of CXCR4 to the degradative endocytic pathway following ligand stimulation by ubiquitinating endosomal sorting complex required for transport ESCRT-0 components HGS and STAM. Targets DTX1 for lysosomal degradation and controls NOTCH1 degradation, in the absence of ligand, through 'Lys-29'-linked polyubiquitination. Ubiquitinates SNX9. Ubiquitinates MAP3K7 through 'Lys-48'-linked conjugation (By similarity). Involved in the regulation of apoptosis and reactive oxygen species levels through the ubiquitination and proteasomal degradation of TXNIP. Mediates the antiapoptotic activity of epidermal growth factor through the ubiquitination and proteasomal degradation of p15 BID. Ubiquitinates BRAT1 and this ubiquitination is enhanced in the presence of NDFIP1.

NCBI and Uniprot Product Information

• NCBI GI #: 380420335
• NCBI GeneID: 83737
• NCBI Accession #: NP_001244066.1
• NCBI GenBank Nucleotide #: NM_001257137.1
• UniProt Accession #: Q96J02; O43584; Q5QP37; Q5TEL0; Q96F66; Q9BY75; Q9H451; Q9H4U5; A6NEW4; B4E234; E1P5P3; F5H217
• Molecular Weight: 102,803 Da
• NCBI Official Full Name: E3 ubiquitin-protein ligase Itchy homolog isoform 1
• NCBI Official Synonym Full Names: itchy E3 ubiquitin protein ligase
• NCBI Official Symbol: ITCH
• NCBI Official Synonym Symbols: AIF4; AIP4; NAPP1; dJ468O1.1
• NCBI Protein Information: E3 ubiquitin-protein ligase Itchy homolog; NFE2-associated polypeptide 1; atrophin-1 interacting protein 4; itchy E3 ubiquitin protein ligase homolog; dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4))
• UniProt Protein Name: E3 ubiquitin-protein ligase Itchy homolog
• Protein Family: E3 ubiquitin-protein ligase
• UniProt Gene Name: ITCH
• UniProt Synonym Gene Names: Itch; AIP4; NAPP1
• UniProt Entry Name: ITCH_HUMAN

NCBI Description

This gene encodes a member of the Nedd4 family of HECT domain E3 ubiquitin ligases. HECT domain E3 ubiquitin ligases transfer ubiquitin from E2 ubiquitin-conjugating enzymes to protein substrates, thus targeting specific proteins for lysosomal degradation. The encoded protein plays a role in multiple cellular processes including erythroid and lymphoid cell differentiation and the regulation of immune responses. Mutations in this gene are a cause of syndromic multisystem autoimmune disease. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Mar 2012]

Uniprot Description

ITCH: 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. It is involved in the control of inflammatory signaling pathways. 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. 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 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NFKB1. Ubiquitinates RIPK2 by 'Lys-63'-linked conjugation and influences NOD2-dependent signal transduction pathways. Regulates the transcriptional activity of several transcription factors, and probably plays an important role in the regulation of immune response. Ubiquitinates NFE2 by 'Lys-63' linkages and is implicated in the control of the development of hematopoietic lineages. Critical regulator of T-helper (TH2) cytokine development through its ability to induce JUNB ubiquitination and degradation. Ubiquitinates SNX9. Ubiquitinates CXCR4 and HGS/HRS and regulates sorting of CXCR4 to the degradative pathway. It is involved in the negative regulation of MAVS-dependent cellular antiviral responses. Ubiquitinates MAVS through 'Lys-48'-linked conjugation resulting in MAVS proteasomal degradation. Involved in the regulation of apoptosis and reactive oxygen species levels through the ubiquitination and proteasomal degradation of TXNIP. Mediates the antiapoptotic activity of epidermal growth factor through the ubiquitination and proteasomal degradation of p15 BID. Targets DTX1 for lysosomal degradation and controls NOTCH1 degradation, in the absence of ligand, through 'Lys-29'-linked polyubiquitination. Monomer Interacts (via its WW domains) with OCNL, NOTCH1 AND JUN. Interacts with JUNB; the interaction promotes ITCH-mediated ubiquitination and degradation of JUNB. Interacts with NDFIP1 and NDFIP2; this interaction activates the E3 ubiquitin-protein ligase and may induce its recruitment to exosomes. Interacts with ARHGEF7. Interacts with RNF11. Interacts (via the WW 1 domain) with NFE2 (via the PXY motif 1); the interaction promotes 'Lys-63'-linked ubiquitination of NFE2, retains it in the cytoplasm and prevents its transactivation activity. Interacts with FYN; the interaction phosphorylates ITCH on Tyr-420 decreasing binding of JUNB. Interacts (via WW domains) with CXCR4 (via C-terminus); the interaction depends on CXCR4 phosphorylation. Interacts (via WW domains) with PCBP2 within a complex containing ITCH, MAVS and PCBP2. Interacts (via WW domains) with TXNIP (via C-terminus). Interacts with ERBB4, DTX1, SPG20, SNX9 and SNX18. Interacts (via its WW domains) with ATN1. Interacts with Epstein-Barr virus LMP2A. Interacts (via WW domains) with SGK3. Widely expressed. 2 isoforms of the human protein are produced by alternative splicing.

Protein type: Ubiquitin ligase; Ubiquitin conjugating system; EC 6.3.2.19; Ligase; EC 6.3.2.-

Chromosomal Location of Human Ortholog: 20q11.22

Cellular Component: membrane; cytoplasm; plasma membrane; cytoplasmic vesicle; cell cortex; cytosol; nucleus

Molecular Function: ribonucleoprotein binding; protein binding; CXCR chemokine receptor binding; ubiquitin-protein ligase activity; ligase activity

Biological Process: ubiquitin-dependent protein catabolic process; negative regulation of JNK cascade; Notch signaling pathway; entry of virus into host cell; apoptosis; protein ubiquitination during ubiquitin-dependent protein catabolic process; protein ubiquitination; negative regulation of defense response to virus; inhibition of NF-kappaB transcription factor; positive regulation of protein catabolic process; innate immune response; negative regulation of alpha-beta T cell proliferation; regulation of cell growth; inflammatory response; positive regulation of T cell anergy; negative regulation of interferon type I production; defense response to virus; negative regulation of apoptosis

Disease: Autoimmune Disease, Multisystem, With Facial Dysmorphism