概述

  • 产品名称Anti-XLF抗体
    参阅全部 XLF 一抗
  • 描述
    兔多克隆抗体to XLF
  • 经测试应用适用于: IP, IHC-P, ICC/IF, WBmore details
  • 种属反应性
    与反应: Human
    预测可用于: Dog
  • 免疫原

    Synthetic peptide corresponding to Human XLF aa 250 to the C-terminus (C terminal).
    (Peptide available as ab27783)

  • 阳性对照
    • HeLa whole cell or nuclear extract, A431 and Jurkat whole cell extracts.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS. pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab33499 in the following tested applications.

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

应用 Ab评论 说明
IP Use at an assay dependent concentration.
IHC-P 1/125. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).

靶标

  • 功能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.
  • 组织特异性Ubiquitously expressed.
  • 疾病相关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).
  • 序列相似性Belongs to the XLF family.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-XLF antibody 图像



  • Predicted band size : 35 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: XLF knockout HAP1 cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green – ab33499 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab33499 was shown to specifically react with XLF when XLF knockout samples were used. Wild-type and XLF knockout samples were subjected to SDS-PAGE. ab33499 and ab18058 (loading control to Vinculin) were diluted at 1 μg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-XLF antibody (ab33499) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 35 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)

    See Abreview

  • XLF was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to XLF and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, A431 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab33499.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: Bands: 35kDa: XLF; non specific - 37kDa: We are unsure as to the identity of this extra band.
  • ab33499 (1/200) staining XLF in assynchronous, bleomycin treated, human RPE-1 cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (blue). Please refer to abreview for further experimental details.

    See Abreview

  • ICC/IF image of ab33499 stained human HEK 293 cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33499, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • Image courtesy of Human Protein Atlas

    ab33499 staining XLF in human kidney. Paraffin embedded human kidney tissue was incubated with ab33499 (1/125 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab33499 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

Anti-XLF antibody (ab33499)参考文献

This product has been referenced in:
  • Terasawa M  et al. Canonical Non-Homologous End Joining in Mitosis Induces Genome Instability and Is Suppressed by M-phase-Specific Phosphorylation of XRCC4. PLoS Genet 10:e1004563 (2014). WB ; Human . Read more (PubMed: 25166505) »
  • Britton S  et al. A new method for high-resolution imaging of Ku foci to decipher mechanisms of DNA double-strand break repair. J Cell Biol 202:579-95 (2013). WB ; Mouse, Human . Read more (PubMed: 23897892) »

See all 4 Publications for this product

Product Wall

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MRC5VA Lung Epithelial Fibroblast)
Loading amount 15 µg
Specification MRC5VA Lung Epithelial Fibroblast
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Apr 13 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (RPE-1)
Specification RPE-1
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

提交于 Aug 16 2007

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