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AB247678

Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free

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Rabbit Recombinant Monoclonal ITCH/AIP4 antibody. Carrier free. Suitable for WB and reacts with Mouse, Rat, Human samples.

View Alternative Names

E3 ubiquitin-protein ligase Itchy homolog, Itch, Atrophin-1-interacting protein 4, HECT-type E3 ubiquitin transferase Itchy homolog, NFE2-associated polypeptide 1, AIP4, NAPP1, ITCH

3 Images
Western blot - Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free (AB247678)
  • WB

Lab

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free (AB247678)

This data was developed using the same antibody clone in a different buffer formulation (ab108515).

Lanes 1-4 : Merged signal (red and green). Green - ab108515 observed at 103 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108515 Anti-ITCH/AIP4 antibody [EPR4936] was shown to specifically react with ITCH/AIP4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265338 (knockout cell lysate ab258014) was used. Wild-type and ITCH/AIP4 knockout samples were subjected to SDS-PAGE. ab108515 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] (<a href='/en-us/products/primary-antibodies/itch-aip4-antibody-epr4936-ab108515'>ab108515</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ITCH knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ITCH (AIP4) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-itch-aip4-knockout-hela-cell-line-ab265338'>ab265338</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

K-562 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 103 kDa,43 kDa

Observed band size: 103 kDa,57 kDa

false

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free (AB247678)
  • WB

Lab

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free (AB247678)

This data was developed using ab108515, the same antibody clone in a different buffer formulation.

ab108515 was shown to react with ITCH in wild-type   cells in Western blot with loss of signal observed in a ITCH knockout cell line. Wild-type   and ITCH knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab108515 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] (<a href='/en-us/products/primary-antibodies/itch-aip4-antibody-epr4936-ab108515'>ab108515</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 100 µg

Lane 2:

ITCH knock-out HeLa  lysate at 100 µg

false

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free (AB247678)
  • WB

Lab

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] - BSA and Azide free (AB247678)

This data was developed using ab108515, the same antibody clone in a different buffer formulation.

Blocking buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-ITCH/AIP4 antibody [EPR4936] (<a href='/en-us/products/primary-antibodies/itch-aip4-antibody-epr4936-ab108515'>ab108515</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Rat brain lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 103 kDa

Observed band size: 103 kDa

false

Exposure time: 1min

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR4936

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Primary Antibodies

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Reactivity data

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Product details

ab247678 is the carrier-free version of ab108515.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

ITCH also known as AIP4 is an E3 ubiquitin ligase with a molecular mass of approximately 108 kDa. The protein primarily functions in the ubiquitination process which regulates protein degradation via the proteasome. ITCH is known for its role in the post-translational modification of proteins through ubiquitin tagging facilitating the promotion or suppression of signal transduction pathways. Expression of ITCH occurs in various tissues including lymphocytes and epithelial cells indicating its diverse functional roles in different physiological contexts.
Biological function summary

ITCH/AIP4 is integral in cellular regulatory mechanisms. It forms part of a sophisticated complex involved in immune response modulation cell cycle regulation and apoptosis. ITCH influences T-cell receptor signaling by controlling the degradation of proteins involved in these critical pathways. Through the ubiquitination cascade ITCH maintains cellular homeostasis participating in the intricate balance of protein synthesis and degradation necessary for normal cellular function.

Pathways

ITCH/AIP4 plays essential roles in the T-cell receptor and Wnt signaling pathways. It interacts with other proteins such as NEDD4 and CBL which are also E3 ubiquitin ligases to modulate signaling cascades involved in immune responses and cellular growth. In the Wnt pathway ITCH regulates the stability of Dishevelled proteins thereby affecting cellular proliferation and differentiation processes. This involvement in multiple pathways highlights its regulatory versatility and importance in maintaining cellular balance.

ITCH/AIP4 is associated with autoimmune diseases and cancer. Its dysfunction can lead to abnormal immune responses contributing to conditions such as systemic lupus erythematosus. Moreover the protein's role in oncogenesis is linked to its ability to modify tumor suppressor and oncoprotein levels including p53 and c-Jun. Disruption in ITCH's function can result in unchecked cell growth and survival emphasizing its potential as a therapeutic target in cancer treatment.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Acts as an 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 (PubMed : 11046148, PubMed : 14602072, PubMed : 15051726, PubMed : 16387660, PubMed : 17028573, PubMed : 18718448, PubMed : 18718449, PubMed : 19116316, PubMed : 19592251, PubMed : 19881509, PubMed : 20068034, PubMed : 20392206, PubMed : 20491914, PubMed : 23146885, PubMed : 24790097, PubMed : 25631046). Catalyzes 'Lys-29'-, 'Lys-48'- and 'Lys-63'-linked ubiquitin conjugation (PubMed : 17028573, PubMed : 18718448, PubMed : 19131965, PubMed : 19881509). Involved in the control of inflammatory signaling pathways (PubMed : 19131965). Essential component of a ubiquitin-editing protein complex, comprising also TNFAIP3, TAX1BP1 and RNF11, that ensures the transient nature of inflammatory signaling pathways (PubMed : 19131965). Promotes the association of the complex after TNF stimulation (PubMed : 19131965). Once the complex is formed, TNFAIP3 deubiquitinates 'Lys-63' polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains (PubMed : 19131965). This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NFKB1 (PubMed : 19131965). Ubiquitinates RIPK2 by 'Lys-63'-linked conjugation and influences NOD2-dependent signal transduction pathways (PubMed : 19592251). Regulates the transcriptional activity of several transcription factors, and probably plays an important role in the regulation of immune response (PubMed : 18718448, PubMed : 20491914). Ubiquitinates NFE2 by 'Lys-63' linkages and is implicated in the control of the development of hematopoietic lineages (PubMed : 18718448). Mediates JUN ubiquitination and degradation (By similarity). Mediates JUNB ubiquitination and degradation (PubMed : 16387660). 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 (PubMed : 19881509). Ubiquitinates MAVS through 'Lys-48'-linked conjugation resulting in MAVS proteasomal degradation (PubMed : 19881509). Following ligand stimulation, regulates sorting of Wnt receptor FZD4 to the degradative endocytic pathway probably by modulating PI42KA activity (PubMed : 23146885). Ubiquitinates PI4K2A and negatively regulates its catalytic activity (PubMed : 23146885). 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 (PubMed : 14602072, PubMed : 23146885, PubMed : 34927784). Targets DTX1 for lysosomal degradation and controls NOTCH1 degradation, in the absence of ligand, through 'Lys-29'-linked polyubiquitination (PubMed : 17028573, PubMed : 18628966, PubMed : 23886940). Ubiquitinates SNX9 (PubMed : 20491914). Ubiquitinates MAP3K7 through 'Lys-48'-linked conjugation (By similarity). Together with UBR5, involved in the regulation of apoptosis and reactive oxygen species levels through the ubiquitination and proteasomal degradation of TXNIP : catalyzes 'Lys-48'-/'Lys-63'-branched ubiquitination of TXNIP (PubMed : 20068034, PubMed : 29378950). ITCH synthesizes 'Lys-63'-linked chains, while UBR5 is branching multiple 'Lys-48'-linked chains of substrate initially modified (PubMed : 29378950). Mediates the antiapoptotic activity of epidermal growth factor through the ubiquitination and proteasomal degradation of p15 BID (PubMed : 20392206). Ubiquitinates BRAT1 and this ubiquitination is enhanced in the presence of NDFIP1 (PubMed : 25631046). Inhibits the replication of influenza A virus (IAV) via ubiquitination of IAV matrix protein 1 (M1) through 'Lys-48'-linked conjugation resulting in M1 proteasomal degradation (PubMed : 30328013). Ubiquitinates NEDD9/HEF1, resulting in proteasomal degradation of NEDD9/HEF1 (PubMed : 15051726).
See full target information E3 ubiquitin-protein ligase Itchy homolog
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Product promise

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