Overview

  • Product name
    Anti-Y14 antibody [4C4]
    See all Y14 primary antibodies
  • Description
    Mouse monoclonal [4C4] to Y14
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, ICC/IF, ELISA, WB, IPmore details
  • Species reactivity
    Reacts with: Human, Xenopus laevis
  • Immunogen

    Full length native protein (purified) (Human).

Properties

Applications

Our Abpromise guarantee covers the use of ab5828 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
Flow Cyt Use 1-2µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/15. PubMed: 17103222
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa). PubMed: 17103222
IP Use at an assay dependent concentration.

Target

  • Function
    Component of a splicing-dependent multiprotein exon junction complex (EJC) deposited at splice junction on mRNAs. The EJC is a dynamic structure consisting of a few core proteins and several more peripheral nuclear and cytoplasmic associated factors that join the complex only transiently either during EJC assembly or during subsequent mRNA metabolism. Core components of the EJC, that remains bound to spliced mRNAs throughout all stages of mRNA metabolism, functions to mark the position of the exon-exon junction in the mature mRNA and thereby influences downstream processes of gene expression including mRNA splicing, nuclear mRNA export, subcellular mRNA localization, translation efficiency and nonsense-mediated mRNA decay (NMD). The heterodimer MAGOH-RBM8A interacts with PYM that function to enhance the translation of EJC-bearing spliced mRNAs by recruiting them to the ribosomal 48S preinitiation complex. Remains associated with mRNAs in the cytoplasm until the mRNAs engage the translation machinery. Its removal from cytoplasmic mRNAs requires translation initiation from EJC-bearing spliced mRNAs. Associates preferentially with mRNAs produced by splicing. Does not interact with pre-mRNAs, introns, or mRNAs produced from intronless cDNAs. Associates with both nuclear mRNAs and newly exported cytoplasmic mRNAs. Complex with MAGOH is a component of the nonsense mediated decay (NMD) pathway.
  • Tissue specificity
    Ubiquitous.
  • Sequence similarities
    Belongs to the RBM8A family.
    Contains 1 RRM (RNA recognition motif) domain.
  • Cellular localization
    Nucleus. Nucleus speckle. Cytoplasm. Nucleocytoplasmic shuttling protein. Travels to the cytoplasm as part of the exon junction complex (EJC) bound to mRNA. Colocalizes with the core EJC, THOC4, NXF1 and UAP56 in the nucleus and nuclear speckles.
  • Information by UniProt
  • Database links
    • Alternative names
      • Binder of OVCA1 1 antibody
      • Binder of OVCA1-1 antibody
      • BOV 1 antibody
      • BOV 1A antibody
      • BOV 1B antibody
      • BOV 1C antibody
      • BOV-1 antibody
      • BOV1 antibody
      • BOV1A antibody
      • BOV1B antibody
      • BOV1C antibody
      • HSPC 114 antibody
      • HSPC114 antibody
      • MDS 014 antibody
      • MDS014 antibody
      • RBM 8 antibody
      • RBM 8A antibody
      • RBM 8B antibody
      • RBM8 antibody
      • rbm8a antibody
      • RBM8A_HUMAN antibody
      • RBM8B antibody
      • Ribonucleoprotein RBM 8 antibody
      • Ribonucleoprotein RBM 8A antibody
      • Ribonucleoprotein RBM8 antibody
      • Ribonucleoprotein RBM8A antibody
      • RNA binding motif protein 8 antibody
      • RNA binding motif protein 8A antibody
      • RNA binding motif protein 8B antibody
      • RNA binding protein 8A antibody
      • RNA binding protein Y14 antibody
      • RNA-binding motif protein 8A antibody
      • RNA-binding protein 8A antibody
      • RNA-binding protein Y14 antibody
      • ZNRP antibody
      • ZRNP 1 antibody
      • ZRNP1 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-Y14 antibody [4C4] (ab5828)
      Immunocytochemistry/ Immunofluorescence - Anti-Y14 antibody [4C4] (ab5828)
      ICC/IF image of ab5828 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5828, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Western blot - Anti-Y14 antibody [4C4] (ab5828)
      Western blot - Anti-Y14 antibody [4C4] (ab5828)
      All lanes : Anti-Y14 antibody [4C4] (ab5828) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
      Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 21 kDa
      Observed band size: 21 kDa
      Additional bands at: 53 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 8 minutes
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Y14 antibody [4C4] (ab5828)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Y14 antibody [4C4] (ab5828)
      IHC image of ab5828 staining in human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5828, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      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.

    • Flow Cytometry - Anti-Y14 antibody [4C4] (ab5828)
      Flow Cytometry - Anti-Y14 antibody [4C4] (ab5828)
      Overlay histogram showing HeLa cells stained with ab5828 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5828, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    References

    This product has been referenced in:
    • van Koningsbruggen S  et al. FRG1P-mediated aggregation of proteins involved in pre-mRNA processing. Chromosoma 116:53-64 (2007). Read more (PubMed: 17103222) »
    • Nojima T  et al. The interaction between cap-binding complex and RNA export factor is required for intronless mRNA export. J Biol Chem 282:15645-51 (2007). Read more (PubMed: 17363367) »

    See all 3 Publications for this product

    Publishing research using ab5828? Please let us know so that we can cite the reference in this datasheet.

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    Western blot abreview for Anti-Y14 antibody [4C4]

     Average
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293E)
    Loading amount
    20 µg
    Specification
    HEK293E
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    Username

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    Verified customer

    Submitted Jun 05 2007

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