53BP1 Antibody

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-904] - Immunofluorescence of 53BP1 Antibody in HeLa cells. Image from verified customer review.

Western Blot: 53BP1 Antibody [NB100-904] - Western blot analysis of untreated U2OS cell lysate blotted with 53BP1 Antibody. The observed molecular weight here is ~220 kDa, and the theoretical molecular weight is 214 kDa.

Immunohistochemistry: 53BP1 Antibody [NB100-904] - Staining of 53BP1 in human renal cancer cells using DAB with hematoxylin counterstain.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-904] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with 53BP1 Antibody conjugated to Alexa Fluor 488 (Catalog #NB100-904AF488) at 10ug/mL for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: 53BP1 Antibody [NB100-904] - An intracellular stain was performed on HeLa cells (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

Western Blot: 53BP1 Antibody [NB100-904] - WRAP53beta plays an important role in recruitment of factors involved in HR and NHEJ to DNA breaks in A2780 cells. A2780 cells were transfected with siControl or siWRAP53#2 oligonucleotides for 48 h; exposed to IR (6 Gy), and 1 h later immunostained for WRAP53beta, gamma-H2AX, MDC1, RNF168, 53BP1, RAD51 and beta-actin. We could not assess the protein levels of BRCA1 due to a lack of antibodies that work for western blotting. The observed molecular weight is 275 kDa and the theoretical molecular weight for the whole endogenous protein is 214 kDa. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015250) licensed under a CC-BY licence.

Flow Cytometry: 53BP1 Antibody [NB100-904] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-904C) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-904] - Immunofluorescence of 53BP1 Antibody in C33A cells. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-904] - Immunofluorescence of 53BP1 in HeLa cells with FITC (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Flow Cytometry: 53BP1 Antibody [NB100-904] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-904AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

• Reactivity: Hu, Mu, Other
• Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP, ISH, PLA, KD
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

53BP1 Antibody Summary

• Immunogen: 53BP1 Antibody was made to a synthetic peptide corresponding to a portion of human 53BP1 encoded in exon 11, 12 and 19 [Uniprot: Q12888]. This antibody is a cocktail hybrid of NB100-304 and NB100-305.
• Marker: DNA Double Strand Break Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TP53BP1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000 - 1:10000
  • Flow Cytometry 2 - 5 ug/mL
  • Immunocytochemistry/Immunofluorescence 1:100 - 1:1000
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Paraffin 1:400
  • Immunoprecipitation
  • In-situ Hybridization
  • Proximity Ligation Assay
  • Knockdown Validated
• Application Notes: In Western blot a band was observed at ~220 kDa. Irradiated lysates demonstrate a gel shift. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Use in immunoprecipitation reported in scientific literature (PMID: 25512560). Use in proximity ligation assay reported in scientific literature (PMID: 26734725). Use in In-situ Hybridization reported in scientific literature (PMID: 25407517). Knockdown data (PMID: 31635359).
• Control:

  Human Kidney Whole Tissue Lysate (Adult Whole Tumor)
  Human Salivary Gland Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Human has been tested in WB, IHC and ICC/IF, mouse has only been tested in ICC/IF. Other reactivity reported in scientific literature (PMID: 31105868). Predicted cross-reactivity based on sequence identity: Human (100%), Chimpanzee (99%), Orangutan (99%), Gibbon (98%), Gorilla (98%), Marmoset (95%)

Packaging, Storage & Formulations

• Storage: Store at -20 °C.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for 53BP1 Antibody

• 53BP1
• p202
• p53-binding protein 1
• p53bp1
• TDRD30
• TP53-Binding Protein 1
• TP53BP1
• tumor protein 53-binding protein, 1
• tumor protein p53 binding protein 1
• tumor protein p53-binding protein, 1
• tumor suppressor p53-binding protein 1

Background

Tumor protein p53 binding protein 1 (P53-binding protein 1 or 53BP1) plays a critical role in tumor suppression and is a putative substrate of ATM kinase with a theoretical molecular weight of 214 kDa. Upon DNA damage, it is phosphorylated and relocalizes to the presumptive sites of damage, specifically, double-strand breaks. 53BP1 plays a key role in response to DNA damage, acts as a signaling checkpoint during mitosis, and enhances TP53-mediated transcriptional activation. Originally identified as p53's transcriptional enhancing partner, 53BP1 is known as a key substrate for ataxia telangiectasia mutated (ATM) signaling, whose function to generate gamma H2AX may be partially compensated by the activity of DNA-dependent kinase (DNA-PK). 53BP1 relocalizes to discrete foci overlapping with gamma phosphorylated histone H2AX; demarcating DNA double strand breaks (DSBs) sites following exposure to radiation (1). 53BP1 functions downstream of gamma H2AX-dependent proteins that collectively establish ionizing radiation induced foci at DSBs. 53BP1 is downstream of Mre11/Rad50/NBS1 (MRN complex), MDC1, RNF8, RNF168 and HERC2 which recruit 53BP1 to the DSB site, suggesting a role in DNA repair through genomic stability maintenance (2).

References

1.Henry, E., Souissi-Sahraoui, I., Deynoux, M., Lefevre, A., Barroca, V., Campalans, A.,... Arcangeli, M. L. (2019). Human hematopoietic stem/progenitor cells display ROS-dependent long-term hematopoietic defects after exposure to low dose of ionizing radiations. Haematologica. doi:10.3324/haematol.2019.226936

2.Janoshazi, A. K., Horton, J. K., Zhao, M. L., Prasad, R., Scappini, E. L., Tucker, C. J., & Wilson, S. H. (2020). Shining light on the response to repair intermediates in DNA of living cells. DNA Repair (Amst), 85, 102749. doi:10.1016/j.dnarep.2019.102749

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for 53BP1 Antibody (NB100-904)(90)

Publications using NB100-904

• Bergstrand S, Bohm S, Malmgren H et al. Biallelic mutations in WRAP53 result in dysfunctional telomeres, Cajal bodies and DNA repair, thereby causing Hoyeraal-Hreidarsson syndrome Cell Death Dis Apr 17 2020 [PMID: 32303682] (ICC/IF, Human)
• O'Dea R, Santocanale C Non-canonical regulation of homologous recombination DNA repair by the USP9X deubiquitinase J. Cell. Sci. Jan 21 2020 [PMID: 31964704] (WB)
• Jin S, Cordes N ATM controls DNA repair and mitochondria transfer between neighboring cells Cell Commun. Signal Nov 8 2019 [PMID: 31703695] (WB, ICC/IF, Human)
• Zhang W, Liao CY, Chtatou H et al. Apalutamide Sensitizes Prostate Cancer to Ionizing Radiation via Inhibition of Non-Homologous End-Joining DNA Repair Cancers (Basel) 2019 Oct 18 [PMID: 31635359] (ICC/IF, KD, Human)
• Cebria-Costa JP, Pascual-Reguant L, Gonzalez-Perez A et al. LOXL2-mediated H3K4 oxidation reduces chromatin accessibility in triple-negative breast cancer cells Oncogene Aug 28 2019 [PMID: 31462706] (WB, Human)
• He B, Gnawali N, Hinman AW, et al. Chromosomes missegregated into micronuclei contribute to chromosomal instability by missegregating at the next division Oncotarget Apr 12 2019 [PMID: 31105868] (ICC/IF, Other)
• Yost KE, Clatterbuck Soper SF, Walker RL et al. Rapid and reversible suppression of ALT by DAXX in osteosarcoma cells Sci Rep Mar 14 2019 [PMID: 30872698] (ICC/IF, Human)
• Shine E, Xue M, Patel J et al. Model colibactins exhibit human cell genotoxicity in the absence of host bacteria bioRxiv Sep 30 2018 [PMID: 30403848] (ICC/IF, Human)
• Chakraborty P, Huang JTJ, Hiom K. DHX9 helicase promotes R-loop formation in cells with impaired RNA splicing Nat Commun Oct 19 2018 [PMID: 30341290] (ICC/IF, WB, IP, Human)
• Bayley R, Blakemore D, Cancian L et al. MYBL2 supports DNA double strand break repair in haematopoietic stem cells Cancer Res. Aug 6 2018 [PMID: 30082276] (Human)
• Kang YJ, Yan CT. Regulation of DNA repair in the absence of classical non-homologous end joining DNA Repair (Amst.) Jun 12 2018 [PMID: 29929045] (ICC/IF)
• Bueno MTD, Baldascini M, Richard S, Lowndes NF. Recruitment of lysine demethylase 2A to DNA double strand breaks and its interaction with 53BP1 ensures genome stability. Oncotarget Mar 23 2018 [PMID: 29662616] (ICC/IF, Human)
• Borude P, Bhushan B, Apte U. DNA Damage Response Regulates Initiation of Liver Regeneration Following Acetaminophen Overdose Gene Expr. May 18 2018 [PMID: 29540258] (WB, Mouse)
• Biswas U, Stevense M, Jessberger R. SMC1a Substitutes for Many Meiotic Functions of SMC1b but Cannot Protect Telomeres from Damage Curr. Biol. 2018 Jan 22 [PMID: 29337080] (ICC/IF)
• Hassan S, Esch A, Liby T et al. Pathway-enriched gene signature associated with 53BP1 response to PARP inhibition in triple-negative breast cancer. Mol. Cancer Ther. 2017 Sep 27 [PMID: 28958991] (Human)
• Bao X, Siprashvili Z, Zarnegar BJ et al. CSNK1a1 Regulates PRMT1 to Maintain the Progenitor State in Self-Renewing Somatic Tissue Dev. Cell 2017 Sep 18 [PMID: 28943242] (IHC, Mouse)
• Sivanand S, Rhoades S, Jiang Q et al. Nuclear Acetyl-CoA Production by ACLY Promotes Homologous Recombination Mol. Cell 2017 Jul 20 [PMID: 28689661]
• Foskolou IP, Jorgensen C, Leszczynska KB et al. Ribonucleotide Reductase Requires Subunit Switching in Hypoxia to Maintain DNA Replication. Mol. Cell. Apr 20 2017 [PMID: 28416140] (WB, ICC/IF, Human)
• Ho TT, Warr MR, Adelman ER et al. Autophagy maintains the metabolism and function of young and old stem cells. Nature. Mar 9 2017 [PMID: 28241143]
• Li S, Shu FJ, Li Z et al. Cell-type specific role of the RNA-binding protein, NONO, in the DNA double-strand break response in the mouse testes. DNA Repair (Amst) 2017 Mar [PMID: 28209515] (Mouse)
• Cheng LC, Tan VM, Ganesan S, Drake JM. Integrating phosphoproteomics into the clinical management of prostate cancer. Clin Transl Med 2017 Dec 01 [PMID: 28197968] (ICC/IF, Mouse)
• Kang YJ, Balter B, Csizmadia E et al. Contribution of classical end-joining to PTEN inactivation in p53-mediated glioblastoma formation and drug-resistant survival Nat Commun Jan 17 2017 [PMID: 28094268] (ICC/IF, Mouse)
• Demaria Marco, O'leary Monique N, Chang Jianhui et al. Cellular Senescence Promotes Adverse Effects of Chemotherapy and Cancer Relapse. Cancer Discov 2017 [PMID: 27979832] (ICC/IF, Mouse)
• Jaafar L, Li Z, Li S, Dynan WS SFPQ-NONO and XLF function separately and together to promote DNA double-strand break repair via canonical nonhomologous end joining Nucleic Acids Res. 2017 Feb 27 [PMID: 27924002] (ICC/IF, Human)
• Dilley RL, Verma P, Cho NW et al. Break-induced telomere synthesis underlies alternative telomere maintenance. Nature Oct 19 2016 [PMID: 27760120] (WB, Human)
• Moureau S, Luessing J, Harte EC et al. A role for the p53 tumour suppressor in regulating the balance between homologous recombination and non-homologous end joining Open Biol Sep 1 2016 [PMID: 27655732] (ICC/IF, WB, Human)
• Rantala JK, Kwon S, Korkola J, Gray JW Expanding the Diversity of Imaging-Based RNAi Screen Applications Using Cell Spot Microarrays. Microarrays (Basel) 2013 Apr 11 [PMID: 27605183]
• Moreno A, Carrington JT, Albergante L et al. Unreplicated DNA remaining from unperturbed S phases passes through mitosis for resolution in daughter cells. Proc. Natl. Acad. Sci. U.S.A. Aug 11 2016 [PMID: 27516545] (ICC/IF, Human)
• Saredi G, Huang H, Hammond CM et al. H4K20me0 marks post-replicative chromatin and recruits the TONSL-MMS22L DNA repair complex. Nature Jun 22 2016 [PMID: 27338793] (WB)
• Pederiva C, Bohm S, Julner A, Farnebo M. Splicing controls the ubiquitin response during DNA double-strand break repair. Cell Death Differ. Jun 17 2016 [PMID: 27315300] (ICC/IF, WB)
• Surovtseva YV, Jairam V, Salem AF et al. Characterization of Cardiac Glycoside Natural Products as Potent Inhibitors of DNA Double-Strand Break Repair by a Whole-Cell Double Immunofluorescence Assay. J Am Chem Soc 2016 Mar 23 [PMID: 26927829]
• Olley GJ Disease-associated Brd4 mutation-linking chromatin binding and the DNA damage response Thesis Jul 9 2019
• Rassoolzadeh H, Bohm S, Hedstrom E et al. Overexpression of the scaffold WD40 protein WRAP53b enhances the repair of and cell survival from DNA double-strand breaks. Cell Death Dis Jun 17 2016 [PMID: 27310875] (ICC/IF, WB)
• Avila AI, Illing A, Becker F et al. Xpg limits the expansion of haematopoietic stem and progenitor cells after ionising radiation. Nucleic Acids Res. May 2 2016 [PMID: 27137888] (ICC/IF, Mouse)
• Byrd PJ, Stewart GS, Smith A et al. A Hypomorphic PALB2 Allele Gives Rise to an Unusual Form of FA-N Associated with Lymphoid Tumour Development. PLoS Genet 2016 Mar [PMID: 26990772] (WB)
• Mackenzie KJ, Carroll P, Lettice L et al. Ribonuclease H2 mutations induce a cGAS/STING-dependent innate immune response. EMBO J. 2016 Feb 22 [PMID: 26903602] (ICC/IF, Mouse)
• Rassoolzadeh H, Coucoravas C, Farnebo M. The proximity ligation assay reveals that at DNA double-strand breaks WRAP53B associates with gammaH2AX and controls interactions between RNF8 and MDC1. Nucleus. 2015 Sep 03 [PMID: 26734725] (PLA, Human)
• Park J, Welner RS, Chan MY et al. The DNA Ligase IV Syndrome R278H Mutation Impairs B Lymphopoiesis via Error-Prone Nonhomologous End-Joining. J. Immunol. 2016 Jan 01 [PMID: 26608917] (ICC/IF)
• Hausmann C, Temme A, Cordes N, Eke I. ILKAP, ILK and PINCH1 control cell survival of p53-wildtype glioblastoma cells after irradiation. Oncotarget 2015 Oct 27 [PMID: 26460618]
• Hedstrom E, Pederiva C, Farnebo J et al. Downregulation of the cancer susceptibility protein WRAP53beta in epithelial ovarian cancer leads to defective DNA repair and poor clinical outcome. Cell Death Dis 2015 [PMID: 26426684] (ICC/IF)
• Shine E, Xue M, Patel J et al. Model colibactins exhibit human cell genotoxicity in the absence of host bacteria Aging and disease Jul 10 1905 (ICC/IF, Human)
• Boysen G, Barbieri CE, Prandi D et al. SPOP mutation leads to genomic instability in prostate cancer. Elife 2015 [PMID: 26374986] (ICC/IF)
• Hartlerode AJ, Morgan MJ, Wu Y et al. Recruitment and activation of the ATM kinase in the absence of DNA-damage sensors. Nat. Struct. Mol. Biol. 2015 Aug 17 [PMID: 26280532] (WB, ICC/IF, Mouse)
• Dickreuter E, Eke I, Krause M et al. Targeting of B1 integrins impairs DNA repair for radiosensitization of head and neck cancer cells Oncogene. 2015 Jun 15 [PMID: 26073085] (ICC/IF, Human)
• Wilson CL, Jurk D, Fullard N et al. NFkB1 is a suppressor of neutrophil-driven hepatocellular carcinoma Nat Commun 2015 Apr 17 [PMID: 25879839] (ICC/IF, Human)
• Singh V, McKinney S, Gerton J Persistent DNA Damage and Senescence in the Placenta Impacts Developmental Outcomes of Embryos SSRN Journal Jan 23 2020 (Human)
• Anbalagan S, Biasoli D, Leszczynska KB et al. In Vitro Radiosensitization of Esophageal Cancer Cells with the Aminopeptidase Inhibitor CHR-2797. Radiat. Res. 2015 Aug 20 [PMID: 26291737] (ICC/IF, Human)
  
  Details:
  53BP1 antibody was used for ICC/IF analysis of FLO-1 cells that were pretreated for 8 hours with DMSO or CHR-2797 followed by exposure to either 0 or 2 Gy radiation (FIG. 3).
• Raymond AA, Benhamouche S, Neaud V et al. Reptin regulates DNA double strand breaks repair in human hepatocellular carcinoma PLoS ONE 2015 Apr 16 [PMID: 25875766] (WB, ICC/IF, Human)
• Kondratova A, Watanabe T, Marotta M et al. Replication fork integrity and intra-S phase checkpoint suppress gene amplification. Nucleic Acids Res 2015 Feb 11 [PMID: 25672394] (ICC/IF)
  
  Details:
  Using the DyLight 488 conjugated version of NB100-904, catalog number NB100-904G.
• Velummailum R. Involvement of NLRP5 in the maintenance of genome integrity in murine oocytes [Internet]. 2011
• Gallina I, Colding C, Henriksen P et al. Cmr1/WDR76 defines a nuclear genotoxic stress body linking genome integrity and protein quality control Nat Commun 2015 Mar 30 [PMID: 25817432] (ICC/IF, Human)
• Luo Z, Feng X, Wang H et al. Mir-23a induces telomere dysfunction and cellular senescence by inhibiting TRF2 expression. Aging Cell. 2015 Mar 06 [PMID: 25753893] (WB, Human)
• Marin-Vicente C, Domingo-Prim J, Eberle AB, Visa N. RRP6/EXOSC10 is required for the repair of DNA double-strand breaks by homologous recombination J. Cell. Sci. 2015 Jan 27 [PMID: 25632158] (ICC/IF, Human)
• Goglia AG, Delsite R, Luz AN et al. Identification of Novel Radiosensitizers in a High-Throughput, Cell-Based Screen for DSB Repair Inhibitors. Mol. Cancer Ther. 2014 Dec 15 [PMID: 25512618] (Human)
• Henriksson S, Rassoolzadeh H, Hedstrom E et al. The scaffold protein WRAP53b orchestrates the ubiquitin response critical for DNA double-strand break repair. Genes Dev. 2014 Dec 15 [PMID: 25512560] (WB, ICC/IF, IP, Human)
• Hosler GA, Davoli T, Mender I et al. A Primary Melanoma and its Asynchronous Metastasis Highlight the Role of BRAF, CDKN2A, and TERT. J. Cutan. Pathol. 2014 Nov 19 [PMID: 25407517]
• Li S, Li Z, Shu FJ et al. Double-strand break repair deficiency in NONO knockout murine embryonic fibroblasts and compensation by spontaneous upregulation of the PSPC1 paralog. Nucleic Acids Res. 2014 Aug 06 [PMID: 25100870] (ICC/IF, Mouse)
• Flach J, Bakker ST, Mohrin M et al. Replication stress is a potent driver of functional decline in ageing haematopoietic stem cells. Nature 2014 Jul 30 [PMID: 25079315] (ICC/IF, Mouse)
  
  Details:
  53BP1 antibody used for ICC-IF in murine haematopoietic stem cells /HSCs - Fixed wth 4% PFA 10 min RT, permeabilized in 0.15% Triton X-100 for 2 min RT, blocked in 1% BSA/PBS ON 4C, primary incubation in 1% BSA/PBS for 1 to 2 h at 37 C. Staining images- Figure 1c, 2a, 3e/h.
• Turner Helen C, Sharma P, Perrier J R et al. The RABiT: high-throughput technology for assessing global DSB repair. Radiat Environ Biophys. 2014 Jan 30 [PMID: 24477408] (Human)
• Kofman AV, Kim J, Park SY et al. microRNA-34a promotes DNA damage and mitotic catastrophe. Cell Cycle 2013 Nov 15 [PMID: 24091633] (ICC/IF, Human)
• Martin OA, Ivashkevich A, Choo S et al. Statistical analysis of kinetics, distribution and co-localisation of DNA repair foci in irradiated cells: Cell cycle effect and implications for prediction of radiosensitivity. DNA Repair (Amst). 2013 Jul 25 [PMID: 23891250]
• Naim V, Wilhelm T, Debatisse M, Rosselli F. ERCC1 and MUS81-EME1 promote sister chromatid separation by processing late replication intermediates at common fragile sites during mitosis. Nat Cell Biol 2013 Jun 30 [PMID: 23811686] (ICC/IF, Human)
• Masumoto K, Tsukimoto M, Kojima S. Role of TRPM2 and TRPV1 cation channels in cellular responses to radiation-induced DNA damage. Biochim Biophys Acta 2013 Feb 28 [PMID: 23458684] (ICC/IF, Human)
• Tang J, Cho NW, Cui G et al. Acetylation limits 53BP1 association with damaged chromatin to promote homologous recombination. Nat Struct Mol Biol 2013 Feb 3 [PMID: 23377543] (ICC/IF, Human)
• Fedor Y, Vignard J, Nicolau-Travers ML et al. From single-strand breaks to double-strand breaks during S-phase: a new mode of action of the Escherichia coliCytolethal Distending Toxin Cell Microbiol 2012 Sep 17 [PMID: 22978660] (ICC/IF, Human)
• Goehe RW, Di X, Sharma K et al. The autophagy-senescence connection in chemotherapy; must tumor cells (self) eat before they sleep? J Pharmacol Exp Ther 2012 Aug 28 [PMID: 22927544] (ICC/IF, Human)
• Hewitt G, Jurk D, Marques FD et al. Telomeres are favoured targets of a persistent DNA damage response in ageing and stress-induced senescence Nat Commun 2012 Feb 28 [PMID: 22426229] (ICC/IF, Human)
• Rantala JK, Makela R, Aaltola AR et al. A cell spot microarray method for production of high density siRNA transfection microarrays. BMC Genomics 2011 Mar 28 [PMID: 21443765] (ICC/IF, Human)
• Coleman KA, Greenberg RA. The BRCA1-RAP80 complex regulates DNA repair mechanism utilization by restricting end resection. J Biol Chem. 2011 Feb 18. [PMID: 21335604]
• Zhang Y-W, Regairaz M, Seiler JA et al. Poly(ADP-ribose) polymerase and XPF-ERCC1 participate in distinct pathways for the repair of topoisomerase I-induced DNA damage in mammalian cells. Nucleic Acids Res. 2011 [PMID: 21227924]
• Monaco-Shawver L, Schwartz L, Tuluc F et al. Substance P inhibits natural killer cell cytotoxicity through the neurokinin-1 receptor. J Leukoc Biol;89(1):113-25. 2011 Jan. [PMID: 20940324]
• Buonomo SB, Wu Y, Ferguson D et al. Mammalian Rif1 contributes to replication stress survival and homology-directed repair. J Cell Biol;187(3):385-98. 2009 Nov 2. [PMID: 19948482]
• Spardy N, Covella K, Cha E et al. Human Papillomavirus 16 E7 Oncoprotein Attenuates DNA Damage Checkpoint Control by Increasing the Proteolytic Turnover of Claspin. Cancer Res 2009:0008-5472 CAN-0009-0925. [PMID: 19706760] (WB, ICC/IF, Human)
• Attwooll CL, Akpinar M, Petrini JHJ. The Mre11 Complex the Response to Dysfunctional Telomeres. Mol Cell Biol 2009:MCB00479-00409. [PMID: 19667076]
• Dickey JS, Baird BJ, Redon CE et al. Intercellular communication of cellular stress monitored by {gamma}-H2AX induction. Carcinogenesis;30(10):1686-1695. 2009 [PMID: 19651821]
• Cadot B, Brunetti M, Coppari S et al. Loss of Histone Deacetylase 4 Causes Segregation Defects during Mitosis of p53-Deficient Human Tumor Cells. Cancer Res 69(15):6074-6082. 2009 [PMID: 19622775]
• Rizzo A, Salvati E, Porru M et al. Stabilization of quadruplex DNA perturbs telomere replication leading to the activation of an ATR-dependent ATM signaling pathway. Nucleic Acids Res :gkp582. 2009 [PMID: 19596811]
• Guirouilh-Barbat J, Zhang Y-W, Pommier Y. Induction of glutathione-dependent DNA double-str breaks by the novel anticancer drug brostallicin. Mol Cancer Ther;8(7):1985-1994. 2009 [PMID: 19584235]
• Yu Z, Chen T, Hebert J et al. A Mouse PRMT1 Null Allele Defines an Essential Role for Arginine Methylation in Genome Maintenance Cell Proliferation. Mol Cell Biol;29(11):2982-2996. 2009 [PMID: 19289494]
• Laszlo A, Fleischer I. Heat-Induced Perturbations of DNA Damage Signaling Pathways are Modulated by Molecular Chaperones. Cancer Res;69(5):2042-2049. 2009 [PMID: 19244134]
• Hsiao SJ, Smith S. Sister telomeres rendered dysfunctional by persistent cohesion are fused by NHEJ. J Cell Biol;184(4):515-26. 2009 Feb 23. [PMID: 19221198]
• Adelman CA, De S, Petrini JHJ. Rad50 is dispensable for the maintenance and viability of postmitotic tissues. Mol Cell Biol;29(2):483-92. 2009 Jan. [PMID: 19001091]
• Palm W, Hockemeyer D, Kibe T et al. Functional dissection of human and mouse POT1 proteins. Mol Cell Biol;29(2):471-82. 2009 Jan. [PMID: 18955498]
• Burdak-Rothkamm S, Rothkamm K, Prise KM. ATM acts downstream of ATR in the DNA damage response signaling of bystander cells. Cancer Res;68(17):7059-65. 2008 Sep 1. [PMID: 18757420]
• Schotta G, Sengupta R, Kubicek S et al. A chromatin-wide transition to H4K20 monomethylation impairs genome integrity and programmed DNA rearrangements in the mouse. Genes Dev;22(15):2048-61. 2008 Aug 1. [PMID: 18676810] (ICC/IF, Mouse)
• Guirouilh-Barbat J, Redon C, Pommier Y. Transcription-coupled DNA double-strand breaks are mediated via the nucleotide excision repair and the Mre11-Rad50-Nbs1 complex. Mol Biol Cell;19(9):3969-81. 2008 Sep. [PMID: 18632984]
• Pennarun G, Granotier C, Hoffschir F et al. Role of ATM in the telomere response to the G-quadruplex ligand 360A. Nucleic Acids Res;36(5):1741-54. 2008 Mar. [PMID: 18263609]
• Toledo, L I, M Murga et al. ATR signaling can drive cells into senescence in the absence of DNA breaks. Genes Dev;22(3):297-302. 2008 Feb 1. [PMID: 18245444] (WB, Mouse)
• Murga, M, I Jaco et al. Global chromatin compaction limits the strength of the DNA damage response. J Cell Biol;178(7):1101-8. 2007 Sep 24. [PMID: 17893239] (WB, Human)
• Yanagawa, T, T Funasaka et al. Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Cancer Res;67(18):8682-9. 2007 Sep 15. [PMID: 17875708]

Reviews for 53BP1 Antibody (NB100-904) (4)

Average Rating: 5

(Based on 4 reviews)

We have 4 reviews tested in 2 species: Human, Mouse.

reviewed by: Anonymous ICC Human 01/20/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: HeLa cells
• Species: Human
• Lot: 012815

Comments

• Comments: Used 1:2000 in 0.5% fish skin gelatin in PBST overnight at 4oC for immunofluorescence. Very good signal to noise ratio.

reviewed by: Anonymous IF Human 11/20/2014

Summary

• Application: Immunofluorescence
• Sample Tested: HeLa cells
• Species: Human
• Lot: 010714

Blocking

• Blocking Details: 2% BSA in PBS, 1 h at RT

Primary Anitbody

• Dilution Ratio: 1:2000 in 2% BSA/PBS, 14h at 4C

Secondary Antibody

• Secondary Description: Alexa Fluor® 488 Goat Anti-Mouse IgG (H+L)
• Secondary Manufacturer Cat#: A-11001 from Life Technologies (by Molecular Probes)
• Secondary Concentration: 0.2 ug/ml

Details

• Detection Notes: Zeiss LSM 780 confocal microscope, x63 oil objective
• Fixation Details: 4% Paraformaldehyde as fixative, permeabilization with 0.2% Triton-X100
• Wash Description: 3x washes in PBS for 5 min

reviewed by: Iris Eke IF Human 08/24/2012

Summary

• Application: Immunofluorescence
• Sample Tested: Human tumor cell lines: A549, FaDu, MiaPaCa2, UTSCC15
• Species: Human
• Lot: 041610

Blocking

• Blocking Details: 1% BSA, 30 min, RT

Primary Anitbody

• Dilution Ratio: 1:2000 1h RT

Secondary Antibody

• Secondary Description: AlexaFluor488 (anti-Rb)
• Secondary Manufacturer Cat#: Invitrogen A11034
• Secondary Concentration: 1:2000

Details

• Detection Notes: images with LSM
• Fixation Details: Fixation: 3%FA, Permeabilisation: Triton-X, Buffer: 1%BSA, 1h incubation RT
• Wash Description: 1xPBS, 6x

reviewed by: Olivia Patsalos IF Mouse 05/22/2012

Summary

• Application: Immunofluorescence
• Sample Tested: MEFs, HeLa, C33A, CaSki
• Species: Mouse
• Lot: 041912
• Comments: Complete protocol can be found at: delangelab.rockefeller.edu/protocols_files/IF_protocol.pdf

Blocking

• Blocking Details: PBG blocking 30mins @RT

Primary Anitbody

• Dilution Ratio: 1:500 2h @RT

Secondary Antibody

• Secondary Description: FITC conjugated Donkey anti-rabbit IgG
• Secondary Manufacturer Cat#: 711-095-152
• Secondary Concentration: 1:250

Details

• Detection Notes: Zeiss Observer A1, Axiovision
• Fixation Details: 4% PFH fixation 10mins @RT, Triton-X permeabilization 10mins @RT, PBS wash 2x5mins @RT
• Wash Description: PBG wash 3x5mins @RT

Comments

• Comments: Complete protocol can be found at: delangelab.rockefeller.edu/protocols_files/IF_protocol.pdf

FAQs for 53BP1 Antibody (NB100-904). (Showing 1 - 5 of 5 FAQs).

1. We're looking for anti53BP1 antibody covalently labeled with a fluorofor preferabley Cy3. Do you make it?
   • We sell 53BP1 antibodies available conjugated to DyLight 488, 550 (very similar in spectrum to Cy3) and 650. You may also purchase the unlabeled antibodies and conjugate them to the fluorophores of your choice. The catalog numbers that you may be interested in are: NB100-904R, NB100-304R, NB100-305R.
2. Hello I want to ask you for an advice in case of antibodies. In our research we focus on DNA repair foci and use your Novus antibodies for phosphorylated histone H2AX (monoclonal mouse) and protein 53BP1(polyclonal rabbit). Now we want to use antobody for phosphorylated 53BP1 protein to increase specifity and I want to ask you which from your NOVUS antibodies will be the best for us.
   • We currently stock just one antibody to phosphorylated 53BP1, which you can see at the following link: NB100-1803. This is guaranteed for detection of 53BP1 (pSer25) in human and mouse samples by WB, FLOW, ICC/IF,IHC-P, IP and PLA. As this antibody is a rabbit polyclonal, you would be unable to stain for both 53BP1 and its phosphorylated isoform in the same sample, unless you used an antibody from a different host for detection of total 53BP1 or used directly conjugated primaries. Our mouse monoclonal to 53BP1, with catalogue number NBP2-25028, is validated for detection of the human protein by WB, FLOW and IHC. Unfortunately we do not currently stock any other antibodies to phosphorylated 53BP1.
3. I would like to know if these antibodies are suitable with IHC techniques using HRP conjugated antibodies and DAB as chromogen? Cat# NB200-175, NB100-904 and NB100-142
   • Yes, all of these antibodies are useful for IHC-P with DAB based detection. Please note that the PTEN Antibody (28H6) NB200-175 has been discontinued, however.
4. I would like to know if this ab would also work for phosphorylated 53BP1? Or if you have another ab that would detect it?
   • NB100-904 is a cocktail of NB100-304 and NB100-305. These two antibodies recognize between residues 350 and 400 and residues 1925 and the C-terminus (residue 1972), respectively. There are multiple phosphorylations between residues 350 and 400 and as you can see from our datasheet, NB100-304 recognizes 53BP1. We do sell one phosphospecific antibody to 53BP1, catalog number NB100-1803.
5. Is there a recommended IHC-P protocol for the 53BP1 Antibody (NB100-904)?
   • We do not have a specific protocol outside of our general protocol for IHC for NB100-904. However, I would ensure that the protocol used includes ample permeabilization for a nuclear signal (usually 0.1-0.5% Triton X in the blocking and diluent buffers or a separate permeabilization step) and heat induced antigen retrieval with pH 6.0 citrate buffer. Our general protocols for both IHC-P and antigen retrieval are linked below. IHC-P Protocol Antigen Retrieival Protocol

Bioinformatics

• Gene Symbol: TP53BP1
• Entrez:
  • 7158(Human)
• Uniprot:
  • Q12888()