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  1. Link

    products/primary-antibodies/53bp1-antibody-epr21732-ab175188.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA Damage & Repair DNA Damage Response p53
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
Reviews (1) Submit a question References (2)

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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

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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

  • 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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR2173(2)
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • p53
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • BRCT Domain Proteins
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • p53 pathway

Associated products

  • Alternative Versions

    • Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Hep G2 whole cell lysate (ab166833)

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.
  • Target information above from: UniProt accession Q12888 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7158 Human
    • Omim: 605230 Human
    • SwissProt: Q12888 Human
    • Unigene: 440968 Human
    • 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)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    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

    Show All Reviews Q&A
    Submit a review Submit a question

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

    Excellent
    Abreviews
    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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