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  1. Link

    53bp1-antibody-epr21722-bsa-and-azide-free-ab222232.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 [EPR2172(2)] - BSA and Azide free (ab222232)

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Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Flow Cytometry (Intracellular) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
  • Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2172(2)] to 53BP1 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Unconjugated

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Anti-53BP1 antibody [EPR2172(2)] (ab175933)

View more associated products

Overview

  • Product name

    Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free
    See all 53BP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR2172(2)] to 53BP1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HepG2 and HeLa cell lysate and human fetal heart and fetal brain tissue lysates, mouse heart and rat heart tissue lysates. IHC-P: human colon, liver carcinoma and tonsil, mouse and rat liver tissues. ICC/IF: HepG2 cells.
  • General notes

    ab222232 is the carrier-free version of ab175933.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2172(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 [EPR2172(2)] (ab175933)
    • Alexa Fluor® 488 Anti-53BP1 antibody [EPR2172(2)] (ab237174)
    • Alexa Fluor® 647 Anti-53BP1 antibody [EPR2172(2)] (ab307631)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

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

    • HeLa whole cell lysate (ab150035)
    • Hep G2 whole cell lysate (ab166833)
    • HeLa whole cell lysate (ab29545)
    • Mouse heart tissue lysate - total protein (0 days) (ab7193)
    • Mouse heart tissue lysate - total protein (14 days) (ab7194)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab222232 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
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 450 kDa (predicted molecular weight: 214 kDa).
Flow Cyt (Intra)
Use at an assay dependent concentration.
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 450 kDa (predicted molecular weight: 214 kDa).
Flow Cyt (Intra)
Use at an assay dependent concentration.

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
    • Entrez Gene: 27223 Mouse
    • Entrez Gene: 296099 Rat
    • Omim: 605230 Human
    • SwissProt: Q12888 Human
    • SwissProt: P70399 Mouse
    • Unigene: 440968 Human
    • Unigene: 383499 Mouse
    • Unigene: 481841 Mouse
    see all
  • 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

  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling 53BP1 with purified ab175933 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/200) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

  • Flow Cytometry (Intracellular) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Flow Cytometry (Intracellular) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    ab175933 staining 53BP1 in the human cell line HepG2 (human hepatocellular carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/30. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody. 

    Isoytype control: Rabbit monoclonal IgG (Black) 

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue) 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933). 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

    Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175933).

  • Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)
    Anti-53BP1 antibody [EPR2172(2)] - BSA and Azide free (ab222232)

Protocols

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

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (0)

Publishing research using ab222232? Please let us know so that we can cite the reference in this datasheet.

ab222232 has not yet been referenced specifically in any publications.

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