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Itchy homolog E3 ubiquitin protein ligase

Antibody: CPTC-ITCH-1
Background

Catalog Number:

CPTC-ITCH-1

RRID:

AB_2617281

Target Antigen:

Itchy homolog E3 ubiquitin protein ligase

Isotype:

IgG2b

Species:

Mouse Monoclonal Antibody

Last Updated:

09/23/2021

Antigen Recognition(s):

Recombinant Full-length, Endogenous

Purchase
External Links
Characterization Data [Compare Characterization Data]
  IHC HPA

CPTC-ITCH-1 Evaluation by the Human Protein Atlas

Result: Negative

This PDF contains the evaluation results provided by the Human Protein Atlas (www.proteinatlas.org)


Characterization SOP Files

  Immunofluorescence - HPA
Click to enlarge image Results provided by the Human Protein Atlas (www.proteinatlas.org). The subcellular location is supported by literature. Immunofluorescent staining of human cell line HeLa shows localization to nucleoplasm & vesicles. Human assay: HeLa fixed with PFA, dilution: 1:2000 Human assay: MCF7 fixed with PFA, dilution: 1:2000 Human assay: U-2 OS fixed with PFA, dilution: 1:2000 Click image to enlarge

CPTC-ITCH-1 Evaluation by the Human Protein Atlas

Result: Positive

Results provided by the Human Protein Atlas (www.proteinatlas.org). The subcellular location is supported by literature. Immunofluorescent staining of human cell line HeLa shows localization to nucleoplasm & vesicles. Human assay: HeLa fixed with PFA, dilution: 1:2000
Human assay: MCF7 fixed with PFA, dilution: 1:2000
Human assay: U-2 OS fixed with PFA, dilution: 1:2000


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA (ie, binding of Antibody to Antigen coated plate). Note: B50% represents the concentration of Ab required to generate 50% of maximum binding. Click image to enlarge

CPTC-ITCH-1 Indirect ELISA

Result: Positive

Indirect ELISA (ie, binding of Antibody to Antigen coated plate). Note: B50% represents the concentration of Ab required to generate 50% of maximum binding.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-ITCH-1 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - HPA - tissue or cell lysate
Click to enlarge image Results provided by the Human Protein Atlas (www.proteinatlas.org). Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. Antibody dilution: 1:500. Click image to enlarge

CPTC-ITCH-1 Evaluation by the Human Protein Atlas

Result: Positive

Results provided by the Human Protein Atlas (www.proteinatlas.org). Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. Antibody dilution: 1:500.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western Blot using CPTC-ITCH-1 as primary Ab against ITCH (rAg 00010) (lane 2). Also included are molecular wt. standards (lane 1) and mouse IgG control (lane 3). Click image to enlarge

CPTC-ITCH-1 Western blot

Result: Positive

Western Blot using CPTC-ITCH-1 as primary Ab against ITCH (rAg 00010) (lane 2). Also included are molecular wt. standards (lane 1) and mouse IgG control (lane 3).


Characterization SOP Files

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Antibody: CPTC-ITCH-2
Background

Catalog Number:

CPTC-ITCH-2

RRID:

AB_2617282

Target Antigen:

Itchy homolog E3 ubiquitin protein ligase

Isotype:

IgG2b

Species:

Mouse Monoclonal Antibody

Last Updated:

09/09/2021

Antigen Recognition(s):

Recombinant Full-length, Endogenous

Purchase
External Links
Characterization Data [Compare Characterization Data]
  IHC HPA

CPTC-ITCH-2 Evaluation by the Human Protein Atlas

Result: Negative

Results provided by the Human Protein Atlas (www.proteinatlas.org)


Characterization SOP Files

  Immunofluorescence - HPA
Click to enlarge image Results provided by the Human Protein Atlas (www.proteinatlas.org). The subcellular location is supported by literature. Immunofluorescent staining of human cell line MCF7 shows localization to nucleoplasm. Human assay: HeLa fixed with PFA, dilution: 1:75 Human assay: MCF7 fixed with PFA, dilution: 1:75 Click image to enlarge

CPTC-ITCH-2 Evaluation by the Human Protein Atlas

Result: Positive

Results provided by the Human Protein Atlas (www.proteinatlas.org). The subcellular location is supported by literature. Immunofluorescent staining of human cell line MCF7 shows localization to nucleoplasm. Human assay: HeLa fixed with PFA, dilution: 1:75
Human assay: MCF7 fixed with PFA, dilution: 1:75


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA (ie, binding of Antibody to Antigen coated plate). Note: B50% represents the concentration of Ab required to generate 50% of maximum binding. Click image to enlarge

CPTC-ITCH-2 Indirect ELISA

Result: Positive

Indirect ELISA (ie, binding of Antibody to Antigen coated plate). Note: B50% represents the concentration of Ab required to generate 50% of maximum binding.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-ITCH-2 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western Blot using CPTC-ITCH-2 as primary Ab against ITCH (rAg 00010) (lane 2). Also included are molecular wt. standards (lane 1) and mouse IgG control (lane 3). Click image to enlarge

CPTC-ITCH-2 Western blot

Result: Positive

Western Blot using CPTC-ITCH-2 as primary Ab against ITCH (rAg 00010) (lane 2). Also included are molecular wt. standards (lane 1) and mouse IgG control (lane 3).


Characterization SOP Files

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Antibody: CPTC-ITCH-3
Background

Catalog Number:

CPTC-ITCH-3

RRID:

AB_2617283

Target Antigen:

Itchy homolog E3 ubiquitin protein ligase

Isotype:

IgG2b

Species:

Mouse Monoclonal Antibody

Last Updated:

08/25/2021

Antigen Recognition(s):

Recombinant Full-length

Purchase
Characterization Data [Compare Characterization Data]
  IHC HPA

CPTC-ITCH-3 Evaluation by the Human Protein Atlas

Result: Negative

Results provided by the Human Protein Atlas (www.proteinatlas.org)


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA (ie, binding of Antibody to Antigen coated plate). Note: B50% represents the concentration of Ab required to generate 50% of maximum binding. Click image to enlarge

CPTC-ITCH-3 Indirect ELISA

Result: Positive

Indirect ELISA (ie, binding of Antibody to Antigen coated plate). Note: B50% represents the concentration of Ab required to generate 50% of maximum binding.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-ITCH-3 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western Blot using CPTC-ITCH-3 as primary Ab against ITCH (rAg 00010) (lane 2). Also included are molecular wt. standards (lane 1) and mouse IgG control (lane 3). Click image to enlarge

CPTC-ITCH-3 Western blot

Result: Positive

Western Blot using CPTC-ITCH-3 as primary Ab against ITCH (rAg 00010) (lane 2). Also included are molecular wt. standards (lane 1) and mouse IgG control (lane 3).


Characterization SOP Files

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Antigen: Itchy homolog E3 ubiquitin protein ligase

Background

NCI Identification Number:

00010

Antigen Name:

Itchy homolog E3 ubiquitin protein ligase

CPTC Name:

CPTC-ITCH

Aliases:

itchy E3 ubiquitin protein ligase; AIP4; NAPP1; NFE2-associated polypeptide 1; itchy (mouse homolog) E3 ubiquitin protein ligase; itchy E3 ubiquitin protein ligase homolog (mouse); AIF4; atrophin-1 interacting protein 4; dJ468O1.1; dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4)); E3 ubiquitin-protein ligase Itchy homolog; itchy E3 ubiquitin protein ligase homolog; EC 6.3.2; Atrophin-1-interacting protein 4

Function:

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.

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. 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 (By similarity). 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

Chromosomal Localization:

20q11.22

Expression System:

E. coli

Accession Number:

NM_008395

UniProt Accession Number:

Q96J02

DNA Source:

Mammalian Gene Collection

Immunogen:

Recombinant Full Length Protein

Vector Name:

pET28-MHL

Extinction Coefficient:

Buffers:

PBS, pH 7.4

Expressed Sequence:

mhhhhhhssgrenlyfqgPSRPPPPTPRRPASVNGSPSATSESDGSSTGS
LPPTNTNTNTSEGATSGLIIPLTISGGSGPRPLNPVTQAPLPPGWEQRVD
QHGRVYYVDHVEKRTTWDRPEPLPPGWERRVDNMGRIYYVDHFTRTTTWQ
RPTLESVRNYEQWQLQRSQLQGAMQQFNQRFIYGNQDLFATSQSKEFDPL
GPLPPGWEKRTDSNGRVYFVNHNTRITQWEDPRSQGQLNEKPLPEGWEMR
FTVDGIPYFVDHNRRTTTYIDPRTGKSALDNGPQIAYVRDFKAKVQYFRF
WCQQLAMPQHIKITVTRKTLFEDSFQQIMSFSPQDLRRRLWVIFPGEEGL
DYGGVAREWFFLLSHEVLNPMYCLFEYAGKDNYCLQINPASYINPDHLKY
FRFIGRFIAMALFHGKFIDTGFSLPFYKRILNKPVGLKDLESIDPEFYNS
LIWVKENNIEECDLEMYFSVDKEILGEIKSHDLKPNGGNILVTEENKEEY
IRMVAEWRLSRGVEEQTQAFFEGFNEILPQQYLQYFDAKELEVLLCGMQE
IDLNDWQRHAIYRHYARTSKQIMWFWQFVKEIDNEKRMRLLQFVTGTCRL
PVGGFADLMGSNGPQKFCIEKVGKENWLPRSHTCFNRLDLPPYKSYEQLK
EKLLFAIEETEGFGQE

Native Sequence:

Calculated Isoelectric Point:

6.4

Molecular Weight:

77784

Last Updated:

08/22/2020

Links

Characterization Data

Gel

Click to enlarge image PAGE of ITCH (rAg 00010) with molecular weight standards in lane 1
Click image to enlarge

CPTC-ITCH PAGE

PAGE of ITCH (rAg 00010) with molecular weight standards in lane 1

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SOPs

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