53BP1 Antibody Pack

Knockout Validated: 53BP1 Antibody Pack [NB100-926] - 53BP1 was detected in immersion fixed HeLa cells (left) but was not detected in 53BP1 knockout Hela cells (right) using Rabbit Anti-human 53BP1 polyclonal antibody (Catalog #NB100-304) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

Western Blot: 53BP1 Antibody Pack [NB100-926] - Western blot analysis of human 53BP1, blotted with 53BP1 antibody (Catalog #NB100-305). Sample: Whole cell lysate (20 ug/lane) from U2Os cells resolved on a 3 to 8% trisacetate gel. A band appears at approximately the theoretical molecular weight of 214 kDa.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody Pack [NB100-926] - Exosomes modulate the repair of DNA DSBs in irradiated recipient cells. Representative images of 53BP1 foci in BHY cells 6 hours after 2 Gy and transfer of BHY exosomes isolated 24 hours after irradiation with 0, 3, 6 or 9 Gy (53BP1 foci green, nuclei blue) using NB100-305. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0152213), licensed under a CC-BY licence.

Immunohistochemistry: 53BP1 Antibody Pack [NB100-926] - Immunohistochemical analysis of human breast tumors stained with 53BP1 antibody (Catalog #NB100-304). Image from verified customer review.

Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-305AF488) (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 Alexa Fluor 488.

Western Blot: 53BP1 Antibody Pack [NB100-926] - Detection of Human [deleted mouse] 53BP1 by Western Blot on Whole cell lysate from U2OS or 293T cells resolved on a 3 to 8% tris-acetate gel using 53BP1 Antibody (Catalog #NB100-304).

Immunohistochemistry: 53BP1 Antibody Pack [NB100-926] - Immunohistochemical staining of placental villi with 53BP1 Antibody (Catalog #NB100-305) at 40X magnification.

Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - Detection of 53BP1. 1 million Jurkat cells were fixed, permeabilized, and stained with 1.5 mcg/ml 53BP1 Antibody (Catalog #NB100-304) in a 150 mcl reaction. Isotype control (black), anti-53bp1 (red).

Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - An intracellular stain was performed on RH-30 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (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 Alexa Fluor 647.

• Reactivity: Hu, Mu
• Applications: WB, Flow, ICC/IF, IHC, KO
• Host: Rabbit
• Concentration: Concentration of individual antibodies may be found on the vial label. If unlisted please contact technical services.

53BP1 Antibody Pack Summary

• Immunogen: See individual datasheets.
• Specificity: The antibodies included in 53BP1 Antibody Pack are specific for 53BP1.
• Host: Rabbit
• Gene: TP53BP1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Knockout Validated
  • Western Blot
• Application Notes: The antibodies included in this pack are useful for Western blot and immunocytochemistry. See individual datasheets for suggested working dilutions.
  See individual datasheets for validated applications.

Reactivity Notes

Human and mouse. Other species have not been tested. Predicted cross-reactivity based on sequence identity: Chimpanzee (99%), Orangutan (99%), Gorilla (98%), Gibbon (98%), Marmoset (95%)

Packaging, Storage & Formulations

• Storage: Store at -20C.
• Buffer: Tris-citrate/phosphate buffer, pH 7 to 8
• Preservative: 0.09% Sodium Azide
• Concentration: Concentration of individual antibodies may be found on the vial label. If unlisted please contact technical services.
• Purity: Immunogen affinity purified

Notes

This pack contains 1 vial each of: NB100-304 (0.1 mL) and NB100-305 (0.1 mL).

Alternate Names for 53BP1 Antibody Pack

• 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. Antibody Packs are guaranteed for 1 year from date of receipt.

Publications for 53BP1 Antibody Pack (NB100-926)(3)

Publications using NB100-926

• Wang Z, Yin H, Zhang Y et al. miR-214-mediated downregulation of RNF8 induces chromosomal instability in ovarian cancer cells. Cell Cycle 2014-01-01 [PMID: 25483088] (WB)
• Sin HS, Barski A, Zhang F et al. RNF8 regulates active epigenetic modifications and escape gene activation from inactive sex chromosomes in post-meiotic spermatids Genes Dev 2012-12-15 [PMID: 23249736] (ICC/IF, IF/IHC, Mouse)
• Kang Y, Lee J-H, Hoan NN et al. Protein Phosphatase 5 Regulates the Function of 53BP1 after Neocarzinostatin-induced DNA Damage. J Biol Chem;284(15):9845-9853. 2009-01-01 [PMID: 19176521]

FAQs for 53BP1 Antibody Pack (NB100-926). (Showing 1 - 2 of 2 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.

Bioinformatics

• Gene Symbol: TP53BP1