Product: ITCH Antibody
Catalog: DF8219
Description: Rabbit polyclonal antibody to ITCH
Application: WB
Reactivity: Human
Prediction: Pig, Bovine, Horse, Rabbit, Dog, Chicken
Mol.Wt.: 103 kDa; 103kD(Calculated).
Uniprot: Q96J02
RRID: AB_2841522

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Product Info

Source:
Rabbit
Application:
WB 1:1000-3000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(83%)
Clonality:
Polyclonal
Specificity:
ITCH Antibody detects endogenous levels of total ITCH.
RRID:
AB_2841522
Cite Format: Affinity Biosciences Cat# DF8219, RRID:AB_2841522.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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; Itchy E3 ubiquitin protein ligase, 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;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q96J02 ITCH_HUMAN:

Widely expressed.

Sequence:
MSDSGSQLGSMGSLTMKSQLQITVISAKLKENKKNWFGPSPYVEVTVDGQSKKTEKCNNTNSPKWKQPLTVIVTPVSKLHFRVWSHQTLKSDVLLGTAALDIYETLKSNNMKLEEVVVTLQLGGDKEPTETIGDLSICLDGLQLESEVVTNGETTCSENGVSLCLPRLECNSAISAHCNLCLPGLSDSPISASRVAGFTGASQNDDGSRSKDETRVSTNGSDDPEDAGAGENRRVSGNNSPSLSNGGFKPSRPPRPSRPPPPTPRRPASVNGSPSATSESDGSSTGSLPPTNTNTNTSEGATSGLIIPLTISGGSGPRPLNPVTQAPLPPGWEQRVDQHGRVYYVDHVEKRTTWDRPEPLPPGWERRVDNMGRIYYVDHFTRTTTWQRPTLESVRNYEQWQLQRSQLQGAMQQFNQRFIYGNQDLFATSQSKEFDPLGPLPPGWEKRTDSNGRVYFVNHNTRITQWEDPRSQGQLNEKPLPEGWEMRFTVDGIPYFVDHNRRTTTYIDPRTGKSALDNGPQIAYVRDFKAKVQYFRFWCQQLAMPQHIKITVTRKTLFEDSFQQIMSFSPQDLRRRLWVIFPGEEGLDYGGVAREWFFLLSHEVLNPMYCLFEYAGKDNYCLQINPASYINPDHLKYFRFIGRFIAMALFHGKFIDTGFSLPFYKRILNKPVGLKDLESIDPEFYNSLIWVKENNIEECDLEMYFSVDKEILGEIKSHDLKPNGGNILVTEENKEEYIRMVAEWRLSRGVEEQTQAFFEGFNEILPQQYLQYFDAKELEVLLCGMQEIDLNDWQRHAIYRHYARTSKQIMWFWQFVKEIDNEKRMRLLQFVTGTCRLPVGGFADLMGSNGPQKFCIEKVGKENWLPRSHTCFNRLDLPPYKSYEQLKEKLLFAIEETEGFGQE

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Dog
100
Rabbit
100
Chicken
83
Sheep
0
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q96J02 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
S6 Phosphorylation
S10 Phosphorylation
K34 Ubiquitination
K53 Methylation
K56 Ubiquitination
K64 Ubiquitination
K66 Ubiquitination
K90 Ubiquitination
K107 Ubiquitination
T214 Phosphorylation
S217 Phosphorylation
T218 Phosphorylation
S221 Phosphorylation
S236 Phosphorylation
S240 Phosphorylation P45983 (MAPK8)
S244 Phosphorylation
S251 Phosphorylation
S257 Phosphorylation
T263 Phosphorylation P45983 (MAPK8)
S273 Phosphorylation P45983 (MAPK8)
Y343 Phosphorylation
Y344 Phosphorylation
K350 Ubiquitination
Y375 Phosphorylation
T385 Phosphorylation Q96BR1 (SGK3)
S393 Phosphorylation
Y397 Phosphorylation
Y420 Phosphorylation P06241 (FYN)
K432 Ubiquitination
K446 Ubiquitination
S450 Phosphorylation Q96BR1 (SGK3) , P17612 (PRKACA)
K478 Ubiquitination
Y495 Phosphorylation
T503 Phosphorylation
T504 Phosphorylation
T505 Phosphorylation
Y506 Phosphorylation
K513 Ubiquitination
Y589 Phosphorylation
K665 Ubiquitination
K670 Ubiquitination
S687 Phosphorylation
K858 Ubiquitination
K861 Ubiquitination
K881 Ubiquitination

Research Backgrounds

Function:

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.

PTMs:

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.

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).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed.

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.

Family&Domains:

The WW domains mediate interaction with PPxY motif-containing proteins.

Research Fields

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

· Genetic Information Processing > Folding, sorting and degradation > Ubiquitin mediated proteolysis.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

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