Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-53BP1 antibody [EPR2173(2)] (ab175188)

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Western blot - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
  • Western blot - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
  • Anti-53BP1 antibody [EPR2173(2)] (ab175188)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2173(2)] to 53BP1
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-53BP1 antibody [EPR2173(2)]
    See all 53BP1 primary antibodies

  • Description

    Rabbit monoclonal [EPR2173(2)] to 53BP1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human 53BP1 aa 900-1000 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q12888

  • Positive control

    • WB: HAP1, HeLa, and HepG2 lysates. Human fetal brain and heart lysates. IHC-P: Human cervical carcinoma and prostate hyperplasia tissue. ICC: HepG2 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab175188 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
1/50000 - 1/200000. Predicted molecular weight: 214 kDa.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
1/100 - 1/250.
Notes
WB
1/50000 - 1/200000. Predicted molecular weight: 214 kDa.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for Flow Cyt or IP.

Target

  • Function

    May have a role in checkpoint signaling during mitosis. Enhances TP53-mediated transcriptional activation. Plays a role in the response to DNA damage.

  • Involvement in disease

    Note=A chromosomal aberration involving TP53BP1 is found in a form of myeloproliferative disorder chronic with eosinophilia. Translocation t(5;15)(q33;q22) with PDGFRB creating a TP53BP1-PDGFRB fusion protein.

  • Sequence similarities

    Contains 2 BRCT domains.

  • Post-translational
    modifications

    Asymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.
    Phosphorylated at basal level in the absence of DNA damage. Hyper-phosphorylated in an ATM-dependent manner in response to DNA damage induced by ionizing radiation. Hyper-phosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation.

  • Cellular localization

    Nucleus. Chromosome > centromere > kinetochore. Associated with kinetochores. Both nuclear and cytoplasmic in some cells. Recruited to sites of DNA damage, such as double stand breaks. Methylation of histone H4 at 'Lys-20' is required for efficient localization to double strand breaks.
  • Information by UniProt

  • Database links

    • Alternative names

      • 53 BP1 antibody
      • 53BP1 antibody
      • FLJ41424 antibody
      • MGC138366 antibody
      • p202 antibody
      • p53 binding protein 1 antibody
      • p53 BP1 antibody
      • p53-binding protein 1 antibody
      • p53BP1 antibody
      • TP53 BP1 antibody
      • TP53B_HUMAN antibody
      • Tp53bp1 antibody
      • TRP53 BP1 antibody
      • Tumor protein 53 binding protein 1 antibody
      • Tumor protein p53 binding protein 1 antibody
      • Tumor suppressor p53 binding protein 1 antibody
      • Tumor suppressor p53-binding protein 1 antibody
      see all

    Images

    • Western blot - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      Western blot - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      All lanes : Anti-53BP1 antibody [EPR2173(2)] (ab175188) at 1/50000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : TP53BP1 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 214 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab175188 observed at 213 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab175188 was shown to recognize 53BP1 in wild-type HAP1 cells as signal was lost at the expected MW in TP53BP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TP53BP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab175188 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/50000 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] (ab175188)

      Immunofluorescence analysis of HepG2 cells labeling 53BP1 with ab175188 at a 1/100 dilution.

    • Western blot - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      Western blot - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      All lanes : Anti-53BP1 antibody [EPR2173(2)] (ab175188) at 1/50000 dilution

      Lane 1 : Human fetal heart tissue lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : Human fetal brain tissue lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 214 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] (ab175188)

      Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling 53BP1 with ab175188 at a 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] (ab175188)

      Immunohistochemical analysis of paraffin-embedded Human prostate hyperplasia tissue labeling 53BP1 with ab175188 at a 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-53BP1 antibody [EPR2173(2)] (ab175188)
      Anti-53BP1 antibody [EPR2173(2)] (ab175188)

    References (2)

    Publishing research using ab175188? Please let us know so that we can cite the reference in this datasheet.

    ab175188 has been referenced in 2 publications.

    • Han J  et al. BRCA2 antagonizes classical and alternative nonhomologous end-joining to prevent gross genomic instability. Nat Commun 8:1470 (2017). WB . PubMed: 29133916
    • Lou J  et al. AUNIP/C1orf135 directs DNA double-strand breaks towards the homologous recombination repair pathway. Nat Commun 8:985 (2017). PubMed: 29042561

    Customer reviews and Q&As

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    Immunocytochemistry/ Immunofluorescence abreview for Anti-53BP1 antibody [EPR2173(2)]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% TritonX100 in PBS
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Kirk Mcmanus

    Verified customer

    Submitted Apr 22 2015

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