Overview

  • Product name
    Anti-CtIP antibody
    See all CtIP primary antibodies
  • Description
    Rabbit polyclonal to CtIP
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human CtIP.

    Read Abcam's proprietary immunogen policy

    (Peptide available as ab38676.)

  • Positive control
    • This antibody gave a positive signal in the following Human lysates: Jurkat Whole Cell HeLa Nuclear HepG2 Whole Cell HepG2 Nuclear F9 Whole Cell (data not shown) Y-79 Nuclear Lysate (data not shown) Caco 2 Whole Cell (data not shown)

Properties

Applications

Our Abpromise guarantee covers the use of ab38016 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 Use a concentration of 1 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 101 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Endonuclease that cooperates with the MRE11-RAD50-NBN (MRN) complex in processing meiotic and mitotic double-strand breaks (DSBs) by ensuring both resection and intrachromosomal association of the broken ends. Functions downstream of the MRN complex and ATM, promotes ATR activation and its recruitment to DSBs in the S/G2 phase facilitating the generation of ssDNA. Component of the BRCA1-RBBP8 complex that regulates CHEK1 activation and controls cell cycle G2/M checkpoints on DNA damage. Promotes microhomology-mediated alternative end joining (A-NHEJ) during class-switch recombination and plays an essential role in chromosomal translocations.
  • Involvement in disease
    Seckel syndrome 2
    Jawad syndrome
    Genetic variability in RBBP8 is noted as a factor in BRCA1-associated breast cancer risk (PubMed:21799032). Exhibits sensitivity to tamoxifen in certain breast cancer cell lines (PubMed:18171986).
  • Sequence similarities
    Belongs to the COM1/SAE2/CtIP family.
  • Domain
    The PXDLS motif binds to a cleft in CtBP proteins.
    The damage-recruitment motif is required for DNA binding and translocation to sites of DNA damage.
  • Post-translational
    modifications
    Acetylated. Deacetylation by SIRT6 upon DNA damage promotes DNA end resection.
    Hyperphosphorylation upon ionizing radiation results in dissociation from BRCA1. Phosphorylation at Thr-847 by CDK1 is essential for the recruitment to DNA and the DNA repair function. Phosphorylated on Ser-327 as cells enter G2 phase. This phosphorylation is required for binding BRCA1 and for the G2/M DNA damage transition checkpoint control.
    Ubiquitinated (PubMed:14654780, PubMed:16818604). Ubiquitination at multiple sites by BRCA1 (via its N-terminal RING domain) does not lead to its proteosomal degradation but instead the ubiquitinated RBBP8 binds to chromatin following DNA damage and may play a role in G2/M checkpoint control (PubMed:16818604). Ubiquitinated by RNF138 at its N-terminus (PubMed:26502057).
  • Cellular localization
    Nucleus. Chromosome. Associates with sites of DNA damage in S/G2 phase (PubMed:10764811). Ubiquitinated RBBP8 binds to chromatin following DNA damage (PubMed:16818604).
  • Information by UniProt
  • Database links
    • Alternative names
      • COM1 antibody
      • COM1_HUMAN antibody
      • CtBP interacting protein antibody
      • CtBP-interacting protein antibody
      • CtIP antibody
      • DNA endonuclease RBBP8 antibody
      • JWDS antibody
      • RB binding protein 8 endonuclease antibody
      • RBBP-8 antibody
      • RBBP8 antibody
      • Retinoblastoma-binding protein 8 antibody
      • Retinoblastoma-interacting protein and myosin-like antibody
      • Rim antibody
      • SAE2 antibody
      • SCKL2 antibody
      • Sporulation in the absence of SPO11 protein 2 homolog antibody
      see all

    Images

    • Western blot - Anti-CtIP antibody (ab38016)
      Western blot - Anti-CtIP antibody (ab38016)
      All lanes : Anti-CtIP antibody (ab38016) at 1 µg/ml

      Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
      Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
      Lane 4 : HepG2 nuclear extract lysate (ab14660)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 101 kDa
      Observed band size: 110 kDa (why is the actual band size different from the predicted?)
      Additional bands at: 25 kDa, 250 kDa. We are unsure as to the identity of these extra bands.

    • Immunocytochemistry/ Immunofluorescence - Anti-CtIP antibody (ab38016)
      Immunocytochemistry/ Immunofluorescence - Anti-CtIP antibody (ab38016)
      ICC/IF image of ab38016 stained MCF-7 cells. The cells were fixed with 100% methanol (5 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 (ab38016 at 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa and HepG2 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 1µg/ml.

    References

    ab38016 has not yet been referenced specifically in any publications.

    Publishing research using ab38016? Please let us know so that we can cite the reference in this datasheet.

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