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Rabbit Recombinant Monoclonal 53BP1 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB and reacts with Human, Mouse samples.

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Images

Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (AB249845), expandable thumbnail
  • Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (AB249845), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (AB249845), expandable thumbnail
  • Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (AB249845), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (AB249845), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFIPFlow CytWB
Human
Tested
Tested
Not recommended
Not recommended
Tested
Mouse
Expected
Tested
Not recommended
Not recommended
Tested

Tested
Tested

Species
Human
Dilution info
-
Notes

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

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human, Mouse
Dilution info
-
Notes

-

Target data

Function

Double-strand break (DSB) repair protein involved in response to DNA damage, telomere dynamics and class-switch recombination (CSR) during antibody genesis (PubMed:12364621, PubMed:17190600, PubMed:21144835, PubMed:22553214, PubMed:23333306, PubMed:27153538, PubMed:28241136, PubMed:31135337, PubMed:37696958). Plays a key role in the repair of double-strand DNA breaks (DSBs) in response to DNA damage by promoting non-homologous end joining (NHEJ)-mediated repair of DSBs and specifically counteracting the function of the homologous recombination (HR) repair protein BRCA1 (PubMed:22553214, PubMed:23333306, PubMed:23727112, PubMed:27153538, PubMed:31135337). In response to DSBs, phosphorylation by ATM promotes interaction with RIF1 and dissociation from NUDT16L1/TIRR, leading to recruitment to DSBs sites (PubMed:28241136). Recruited to DSBs sites by recognizing and binding histone H2A monoubiquitinated at 'Lys-15' (H2AK15Ub) and histone H4 dimethylated at 'Lys-20' (H4K20me2), two histone marks that are present at DSBs sites (PubMed:17190600, PubMed:23760478, PubMed:27153538, PubMed:28241136). Required for immunoglobulin class-switch recombination (CSR) during antibody genesis, a process that involves the generation of DNA DSBs (PubMed:23345425). Participates in the repair and the orientation of the broken DNA ends during CSR (By similarity). In contrast, it is not required for classic NHEJ and V(D)J recombination (By similarity). Promotes NHEJ of dysfunctional telomeres via interaction with PAXIP1 (PubMed:23727112).

Alternative names

TP53-binding protein 1, 53BP1, p53-binding protein 1, p53BP1, TP53BP1

Recommended products

Rabbit Recombinant Monoclonal 53BP1 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB and reacts with Human, Mouse samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR2173(2)
Purification technique
Affinity purification
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab249845 is the carrier-free version of Anti-53BP1 antibody [EPR2173(2)] ab175188.

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

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

53BP1 also known as p53-binding protein 1 is a protein that plays an important role in DNA damage response. The 53BP1 protein has a molecular weight of approximately 213 kDa. It is a nuclear protein and it mainly expresses in cells during the DNA damage repair processes. Researchers commonly utilize immunofluorescence and anti-53BP1 antibodies to detect its presence especially for studying DNA repair mechanisms. 53BP1 proves important for maintenance of genomic stability.

Biological function summary

As a DNA damage response mediator 53BP1 functions to recognize and bind to sites of double-strand breaks in DNA promoting non-homologous end joining (NHEJ). This protein is a component of a larger protein complex that recruits factors necessary for DNA repair. Its interaction with DNA lesions also involves chromatin remodeling aiming to facilitate successful repair of the DNA. 53BP1 works in concert with other proteins such as H2AX and MDC1.

Pathways

Researchers observe 53BP1 in processes such as the DNA damage checkpoint and repair pathways. This protein significantly contributes to the pathways involved in maintaining cell cycle checkpoints. It interacts with proteins like ATM and BRCA1 coordinating the cellular response to DNA damage and ensuring that genomic integrity checks occur properly before cell cycle progression continues. These interactions place 53BP1 at a pivotal position in decision-making between repair pathways.

Associated diseases and disorders

Researchers often focus on 53BP1's involvement in cancer development and therapy resistance due to its role in DNA repair. Alterations or deficiencies in 53BP1 levels can contribute to increased susceptibility to tumor development affecting processes like breast cancer and ovarian cancer through its relationship with BRCA1. The improper function of 53BP1 in DNA repair pathways may lead to an accumulation of genetic mutations driving tumorigenesis and impacting the efficacy of certain cancer treatments.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using Anti-53BP1 antibody [EPR2173(2)] ab175188, the same antibody clone in a different buffer formulation.

    Lanes 1 - 3: Merged signal (red and green). Green - Anti-53BP1 antibody [EPR2173(2)] ab175188 observed at 213 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.

    Anti-53BP1 antibody [EPR2173(2)] 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. Anti-53BP1 antibody [EPR2173(2)] ab175188 and Anti-Vinculin antibody [VIN-54] 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: TP53BP1 knockout HAP1 whole cell lysate at 20 µg

    Lane 3: HeLa whole cell lysate at 20 µg

    Predicted band size: 214 kDa

  • Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-53BP1 antibody [EPR2173(2)] ab175188).

    Lanes 1 - 3: Merged signal (red and green). Green - Anti-53BP1 antibody [EPR2173(2)] ab175188 observed at 213 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.

    Anti-53BP1 antibody [EPR2173(2)] 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. Anti-53BP1 antibody [EPR2173(2)] ab175188 and Anti-Vinculin antibody [VIN-54] 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845) at 1/50000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: TP53BP1 knockout HAP1 whole cell lysate at 20 µg

    Lane 3: HeLa whole cell lysate at 20 µg

    Predicted band size: 214 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using Anti-53BP1 antibody [EPR2173(2)] ab175188, the same antibody clone in a different buffer formulation.

    Immunofluorescence analysis of HepG2 cells labeling 53BP1 with Anti-53BP1 antibody [EPR2173(2)] ab175188 at a 1/100 dilution.

  • Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using Anti-53BP1 antibody [EPR2173(2)] ab175188, the same antibody clone in a different buffer formulation.

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

    Lane 1: Human fetal heart tissue lysate at 10 µg

    Lane 2: HepG2 cell lysate at 10 µg

    Lane 3: Human fetal brain tissue lysate at 10 µg

    Predicted band size: 214 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using Anti-53BP1 antibody [EPR2173(2)] ab175188, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling 53BP1 with Anti-53BP1 antibody [EPR2173(2)] 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)] - BSA and Azide free (ab249845), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using Anti-53BP1 antibody [EPR2173(2)] ab175188, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human prostate hyperplasia tissue labeling 53BP1 with Anti-53BP1 antibody [EPR2173(2)] ab175188 at a 1/50 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Western blot - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    53BP1 Western blot staining using rabbit Anti-53BP1 antibody

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-53BP1 antibody [EPR2173(2)] ab175188).

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Anti-Vinculin antibody [EPR8185] - Loading Control ab129002 was used as a Vinculin loading control at a 1/10000 dilution.

    In Western blot, Anti-53BP1 antibody [EPR2173(2)] ab175188 was shown to bind specifically to Dnmt1. Target of interest was observed mainly at 270 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Dnmt1 knockout cell line (lane 2) .

    All lanes: Western blot - Anti-53BP1 antibody [EPR2173(2)] (Anti-53BP1 antibody [EPR2173(2)] ab175188) at 1/1000 dilution

    Lane 1: Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

    Lane 2: Dnmt1 knockout HAP1 whole cell lysate at 20 µg

    Lane 3: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

    Lane 5: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 6: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 270 kDa

    Exposure time: 70s

  • Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-53BP1 antibody [EPR2173(2)] ab175188).

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling 53BP1 with Anti-53BP1 antibody [EPR2173(2)] ab175188 at 1/500 (0.976 µg/ml) dilution followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at a 1/1000 (2 µg) dilution (Green).

    Confocal image showing nuclear staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at a 1/1000 (2 µg) dilution.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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