Overview

  • Product name

    Anti-XLF antibody [EPR15882-36] - C-terminal
    See all XLF primary antibodies
  • Description

    Rabbit monoclonal [EPR15882-36] to XLF - C-terminal
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human XLF aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9H9Q4

  • Positive control

    • Ramos, Jurkat and HepG2 whole cell lysate (ab7900); Human endometrial adenocarcinoma and Mouse liver tissues; HepG2 and NCCIT cells; Ramos cells.
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR15882-36
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab189917 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 1/60.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/250 - 1/500.
WB 1/10000 - 1/50000. Detects a band of approximately 39 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    DNA repair protein involved in DNA nonhomologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination. May serve as a bridge between XRCC4 and the other NHEJ factors located at DNA ends, or may participate in reconfiguration of the end bound NHEJ factors to allow XRCC4 access to the DNA termini. It may act in concert with XRCC6/XRCC5 (Ku) to stimulate XRCC4-mediated joining of blunt ends and several types of mismatched ends that are noncomplementary or partially complementary.
  • Tissue specificity

    Ubiquitously expressed.
  • Involvement in disease

    Defects in NHEJ1 are the cause of severe combined immunodeficiency due to NHEJ1 deficiency (NHEJ1-SCID) [MIM:611291]; also known as autosomal recessive T cell-negative, B cell-negative, NK cell-positive, severe combined immunodeficiency with microcephaly, growth retardation and sensitivity to ionizing radiation or NHEJ1 syndrome. SCID refers to a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia and low or absent antibody levels. Patients with SCID present in infancy with recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. NHEJ1-SCID is characterized by a profound T- and B-lymphocytopenia associated with increased cellular sensitivity to ionizing radiation, microcephaly and growth retardation. Some patients may manifest SCID with sensitivity to ionizing radiation without microcephaly and mild growth retardation, probably due to hypomorphic NHEJ1 mutations.
    Note=A chromosomal aberration involving NHEJ1 is found in a patient with polymicrogyria. Translocation t(2;7)(q35;p22).
  • Sequence similarities

    Belongs to the XLF family.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cernunnos antibody
    • FLJ12610 antibody
    • NHEJ 1 antibody
    • Nhej1 antibody
    • NHEJ1, S. cerevisiae, homolog of antibody
    • NHEJ1_HUMAN antibody
    • Non homologous end joining factor 1 antibody
    • Non-homologous end-joining factor 1 antibody
    • Nonhomologous end joining factor 1 antibody
    • OTTHUMP00000164168 antibody
    • OTTHUMP00000206275 antibody
    • OTTHUMP00000206279 antibody
    • Protein cernunnos antibody
    • XLF antibody
    • XRCC4 like factor antibody
    • XRCC4-like factor antibody
    see all

Images

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: XLF knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab189917 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab189917 was shown to specifically react with XLF in wild type cells as signal was lost in XLF knockout cells. Wild-type and XLF knockout samples were subjected to SDS-PAGE. ab189917 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometrical analysis of Ramos cells labeling XLF with ab189917 at 1/60 compared to a negative control cell. FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.

  • Immunofluorescent analysis of paraformaldehyde-fixed NCCIT cells labeling XLF with ab189917 at 1/250, Goat anti rabbit IgG (Alexa Fluor® 555) at 1/200 and DAPI staining (blue).

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling XLF with ab189917 at 1/250 with prediluted ImmunoHistoprobe(Ready to use) HRP Polymer for Rabbit IgG as secondary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-XLF antibody [EPR15882-36] - C-terminal (ab189917) at 1/10000 dilution

    Lane 1 : Ramos cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : HepG2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 33 kDa
    Additional bands at: 39 kDa. We are unsure as to the identity of these extra bands.

  • Immunofluorescent analysis of paraformaldehyde-fixed HepG2 cells labeling XLF with ab189917 at 1/250,  Goat anti rabbit IgG (Alexa Fluor® 555) at 1/200 and DAPI staining (blue).

  • Immunohistochemical analysis of paraffin-embedded Human endometrial adenocarcinoma tissue labeling XLF with ab189917 at 1/250 with prediluted ImmunoHistoprobe(Ready to use) HRP Polymer for Rabbit IgG as secondary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Li Z  et al. Impaired DNA double-strand break repair contributes to the age-associated rise of genomic instability in humans. Cell Death Differ 23:1765-1777 (2016). Read more (PubMed: 27391797) »
See 1 Publication for this product

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