Overview

  • Product name
    Anti-USP9x antibody [EPR13809(B)] - N-terminal (HRP)
    See all USP9x primary antibodies
  • Description
    Rabbit monoclonal [EPR13809(B)] to USP9x - N-terminal (HRP)
  • Host species
    Rabbit
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human USP9x aa 1-100 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q93008

  • Positive control
    • WB: Jurkat, HeLa and HepG2 whole cell lysates. IHC-P: FFPE human normal testis tissue sections.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab203471 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/5000. Detects a band of approximately 268 kDa (predicted molecular weight: 292 kDa).
IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Deubiquitinase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. May therefore play an important role regulatory role at the level of protein turnover by preventing degradation of proteins through the removal of conjugated ubiquitin. Essential component of TGF-beta/BMP signaling cascade. Regulates chromosome alignment and segregation in mitosis by regulating the localization of BIRC5/survivin to mitotic centromeres. Specifically hydrolyzes both 'Lys-29'- and 'Lys-33'-linked polyubiquitins chains. Specifically deubiquitinates monoubiquitinated SMAD4, opposing the activity of E3 ubiquitin-protein ligase TRIM33.
  • Tissue specificity
    Widely expressed in embryonic and adult tissues.
  • Sequence similarities
    Belongs to the peptidase C19 family.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Deubiquitinating enzyme FAF X antibody
    • Deubiquitinating enzyme FAF-X antibody
    • DFFRX antibody
    • Drosophila fat facets related X linked antibody
    • FAF antibody
    • Fafl antibody
    • Fam antibody
    • Fat facets homolog antibody
    • Fat facets in mammals antibody
    • Fat facets protein related X linked antibody
    • Fat facets protein related, X-linked antibody
    • Fat facets protein-related antibody
    • hFAM antibody
    • MRX99 antibody
    • Probable ubiquitin carboxyl terminal hydrolase FAF X antibody
    • Probable ubiquitin carboxyl-terminal hydrolase FAF-X antibody
    • Ubiquitin carboxyl-terminal hydrolase FAM antibody
    • Ubiquitin specific peptidase 9 X linked antibody
    • Ubiquitin specific peptidase 9, X-linked antibody
    • Ubiquitin specific processing protease FAF X antibody
    • Ubiquitin specific protease 9 X chromosome antibody
    • Ubiquitin thioesterase FAF X antibody
    • Ubiquitin thiolesterase FAF X antibody
    • Ubiquitin thiolesterase FAF-X antibody
    • Ubiquitin-specific protease 9 antibody
    • Ubiquitin-specific-processing protease FAF-X antibody
    • USP9 (gene name) antibody
    • Usp9x antibody
    • USP9X_HUMAN antibody
    • Uubiquitin specific protease 9, X chromosome (fat facets like Drosophila) antibody
    • X chromosome antibody
    • X-linked antibody
    see all

Images

  • All lanes : Anti-USP9x antibody [EPR13809(B)] - N-terminal (HRP) (ab203471) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : USP9X knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 292 kDa
    Observed band size: 268 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    ab203471 was shown to specifically react with USP9x in wild-type HAP1 cells as signal was lost in USP9X knockout cells. Wild-type and USP9X knockout samples were subjected to SDS-PAGE. Ab203471 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • IHC image of USP9x staining in a section of formalin-fixed paraffin-embedded human normal testis tissue sections*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab203471, 1/200 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-USP9x antibody [EPR13809(B)] - N-terminal (HRP) (ab203471) at 1/5000 dilution

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 292 kDa
    Observed band size: 268 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab203471 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

     

     

     

References

ab203471 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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