Human ITCH (AIP4) knockout HeLa cell lysate
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ITCH KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon7.
View Alternative Names
ADMFD, AIF4, Aip-4, Atrophin-1-interacting protein 4, E3 ubiquitin-protein ligase Itchy homolog, EC 6.3.2, 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, Ubiquitin protein ligase ITCH, dJ468O1.1, dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4)), dJ468O1.1 atrophin 1 interacting protein 4 AIP4
- WB
Lab
Western blot - Human ITCH (AIP4) knockout HeLa cell lysate (AB258014)
Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : ITCH knockout HeLa cell lysate (20 ug)
Lane 3 : HAP1 cell lysate (20 ug)
Lane 4 : K-562 cell lysate (20 ug)
ab108515 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 4oC 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
- Sanger seq
Unknown
Sanger Sequencing - Human ITCH (AIP4) knockout HeLa cell lysate (AB258014)
Homozygous : 1 bp insertion in exon7
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-ITCH/AIP4 antibody [EPR4936] (AB108515)](https://content.abcam.com/products/images/itch-aip4-antibody-epr4936-ab108515--western-blot-img105138.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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