Itch Antibody (PrecisionAbTM Antibody)

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Itch Antibody (PrecisionAb<sup>TM</sup> Antibody) gallery image 1

Western blot analysis of whole cell lysates probed with ITCH antibody followed by detection with HRP conjugated Goat anti Mouse IgG (1/5,000, STAR207P) and visualized on the ChemicDoc™ MP with 117 second exposure. Arrow points to ITCH (molecular weight 102 kDa)

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  • Itch Antibody with Control Lysate
  • Mouse anti Itch
  • Itch Antibody with Control Lysate
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    Mouse anti human ITCH antibody recognizes the E3 ubiquitin-protein ligase Itchy homolog, also known as E3 ligase Itchy homolog, NFE2-associated polypeptide 1, atrophin-1 interacting protein 4, AIP4 or dJ468O1. This antibody has been extensively validated for western blotting using whole cell lysates.
    • Product Type
      PrecisionAb™ Monoclonal
    • Isotype
      IgG2b
    • Format
      Purified
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      VMA00405WBdatasheet pdfdatasheet pdf100 µl
      VMA00405
      VMA00405KWBdatasheet pdfdatasheet pdf datasheet pdf10 Westerns
      VMA00405K
      VMA00405KTWBdatasheet pdfdatasheet pdf datasheet pdf2 Westerns
      VMA00405KT
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      References
      Reviews
      -
      • Mouse anti Human ITCH antibody recognizes ITCH, also known as E3 ligase Itchy homolog, NFE2-associated polypeptide 1, atrophin-1 interacting protein 4, AIP4 or dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4)) and itchy E3 ubiquitin protein ligase homolog.

        The ITCH 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 ITCH are a cause of syndromic multisystem autoimmune disease. Alternatively spliced transcript variants encoding multiple isoforms have been observed for ITCH (provided by RefSeq, Mar 2012).

        Mouse anti Human ITCH antibody detects a band of 102 kDa. The antibody has been extensively validated for western blotting using whole cell lysates. This antibody is available with a positive control lysate, in a smaller trial size, and as a standalone antibody.
      • Target Species
        Human
      • Western Blotting
        Anti ITCH detects a band of approximately 102 kDa in A431 cell lysates
        Anti ITCH detects a band of approximately 102 kDa in A431 cell lysates
        Anti ITCH detects a band of approximately 102 kDa in A431 cell lysates
      • Species Cross-Reactivity
        Target SpeciesCross Reactivity
        Mouseyes
        Ratyes
        N.B. Antibody reactivity and working conditions may vary between species.
      • Product Form
        Purified IgG - liquid
        Purified IgG - liquid
        Purified IgG - liquid
      • Preparation
        Pack Size: 10 Westerns100μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from ascites
        Pack Size: 2 Westerns20μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from ascites
        Mouse monoclonal antibody purified by affinity chromatography on Protein G from ascites
        Pack Size: 10 Westerns100μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from ascites
        Pack Size: 2 Westerns20μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from ascites
      • Buffer Solution
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
      • Preservative Stabilisers
        0.09% Sodium Azide (NaN3)
        0.09% Sodium Azide (NaN3)
        0.09% Sodium Azide (NaN3)
      • Immunogen
        Recombinant protein corresponding to aa 257-903 of human ITCH
      • Instructions for Use
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
      • Lysate Composition
        400μg A431 lysate lyophilized in RIPA buffer
        400μg A431 lysate lyophilized in RIPA buffer
      • Lysate Reconstitution
        - If using DDT reconstitute the lyophilized lysate with 190μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 10μl 2M DTT.
        - If using BME reconstitute the lyophilized lysate with 180μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 20μl BME.
        Heat at 95°C for 5 minutes. For 10 well mini gels load 25μl. For other gel and comb formats please refer to the PrecisionAb western blotting protocol.
        - If using DDT reconstitute the lyophilized lysate with 190μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 10μl 2M DTT.
        - If using BME reconstitute the lyophilized lysate with 180μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 20μl BME.
        Heat at 95°C for 5 minutes. For 10 well mini gels load 25μl. For other gel and comb formats please refer to the PrecisionAb western blotting protocol.
      • Storage
        Antibody: Store undiluted at -20oC, avoiding repeated freeze thaw cycles.

        Lysate: Store lyophilized lysate at -20°C. After reconstitution aliquot and store at -20°C for up to 3 months or at -80°C for longer term storage.
        Store undiluted at -20oC, avoiding repeated freeze thaw cycles.
        Antibody: Store undiluted at -20oC, avoiding repeated freeze thaw cycles.

        Lysate: Store lyophilized lysate at -20°C. After reconstitution aliquot and store at -20°C for up to 3 months or at -80°C for longer term storage.
      • Shelf Life
        As supplied, 12 months from date of despatch
        As supplied, 12 months from date of despatch.
        As supplied, 12 months from date of despatch
      • Acknowledgements
        PrecisionAb™ is a trademark of Bio-Rad Laboratories
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
        PrecisionAb™ is a trademark of Bio-Rad Laboratories
      • Regulatory
        For research purposes only
      • UniProt
      • Entrez Gene
      • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000

      • PrecisionAb antibodies have been extensively validated for the western blot application. The antibody has been validated at the suggested dilution. Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Further optimization may be required dependant on sample type.
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Goat anti Mouse IgG (H/L):HRPSTAR207P2 mlWB
        STAR207P
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Goat anti Mouse IgG (H/L):HRPSTAR207P2 mlWB
        STAR207P
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Goat anti Mouse IgG (H/L):HRPSTAR207P2 mlWB
        STAR207P

        Recommended Negative Isotype Control

          Useful Reagents

            Recommended Positive Controls

              Histology Controls

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