53BP1 Antibody (6B3E10)

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (6B3E10) [NBP2-25028] - Staining of human U-2 OS cells using anti-53BP1 antibody. Bottom images: human U-2 OS cells were treated with EPE to induce DNA-damage and showed increase in nuclear spots after the induction of DNA damage with EPE. Image from verified customer review.

Immunohistochemistry: 53BP1 Antibody (6B3E10) [NBP2-25028] - Analysis of paraffin-embedded colon cancer tissues using 53BP1 Antibody with DAB staining.

Western Blot: 53BP1 Antibody (6B3E10) [NBP2-25028] - Western blot analysis using against HEK293 (1) and 53BP1 (AA: 574-773)-hIgGFc transfected HEK293 (2) cell lysate. The observed molecular weight is ~52 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.

Flow Cytometry: 53BP1 Antibody (6B3E10) [NBP2-25028] - Analysis of HepG2 cells using 53BP1 Antibody (green) and negative control (purple).

Immunohistochemistry: 53BP1 Antibody (6B3E10) [NBP2-25028] - Analysis of paraffin-embedded endometrial cancer tissues using 53BP1 Antibody with DAB staining.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (6B3E10) [NBP2-25028] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti- at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: 53BP1 Antibody (6B3E10) [NBP2-25028] - Western blot analysis using 53BP1 Antibody against human 53BP1 recombinant protein. The expected molecular weight is 47.6 kDa which is demonstrated in this analysis, and the theoretical molecular weight of the whole endogenous protein is 214 kDa.

• Reactivity: Hu
• Applications: WB, Flow, ICC/IF, IHC, IHC-P
• Clone: 6B3E10
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

53BP1 Antibody (6B3E10) Summary

• Immunogen: 53BP1 Antibody (6B3E10) was made to a partial recombinant human 53BP1 (between residues 500-800) expressed in E. coli [Uniprot: Q12888]
• Localization: Nuclear
• Marker: DNA Double Strand Break Marker
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: TP53BP1
• Purity: Ammonium sulfate precipitation

Applications/Dilutions

• Dilutions:
  • Western Blot 1:500 - 1:2000
  • Flow Cytometry 1:200 - 1:400
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:200 - 1:1000
  • Immunohistochemistry-Paraffin 1:200 - 1:1000
• Application Notes: ICC/IF reported in a verified customer review.
• Theoretical MW: 213 kDa.
  Disclaimer note: 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.
• Control:

  Human Colon Transverse Whole Tissue Lysate (Adult Whole Normal)
  Human Uterus Whole Tissue Lysate (Adult Whole Tumor)

Reactivity Notes

Predicted cross-reactivity based on sequence identity: Human (100%), Chimpanzee (99%), Orangutan (99%), Gibbon (98%), Gorilla (98%), Marmoset (95%), Canine (93%), Rabbit (92%), Feline (91%), Equine (91%), Panda (91%), Porcine (91%)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS, 0.5% protein stabilizer
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Ammonium sulfate precipitation

Alternate Names for 53BP1 Antibody (6B3E10)

• 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 (NBP2-25028)(2)

Publications using NBP2-25028

• Pike AM, Strong MA, Ouyang JPT, Greider CW TIN2 Functions with TPP1/POT1 To Stimulate Telomerase Processivity Mol Cell Biol. 2019 Oct 2019 [PMID: 31383750] (ICC/IF, Human)
• Nassour J, Martien S, Martin N, et al. Defective DNA single-strand break repair is responsible for senescence and neoplastic escape of epithelial cells. Nat Commun. 2016 Jan 29 [PMID: 26822533] (ICC/IF, Human)

Review for 53BP1 Antibody (NBP2-25028) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Frida Danielsson ICC Human 12/18/2015

Summary

• Application: Immunocytochemistry
• Sample Tested: U-2 OS cells
• Species: Human
• Lot: A-1

Blocking

• Blocking Details: No blocking

Primary Anitbody

• Dilution Ratio: 1:1000 overnight

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG coupled to Alexa 561

Details

• Detection Notes: Confocal microscopy, Leica SP5
• Fixation Details: paraformaldehyde fixation(4%), permeabilization with Triton-X (0.1%)
• Wash Description: PBS 3 times

Comments

• Comments: Shows a increase in nuclear spots after the induction of DNA damage with etoposide.

FAQs for 53BP1 Antibody (NBP2-25028). (Showing 1 - 3 of 3 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 am curious about the antibody for 53BP1(6B3E10), cat # NBP2-25028. anticipated mw is 213kDa, but the western blot images show around 50 kDa, why is it?
   • The sample on the WB is a partial protein: TP53BP1 (AA: 574-773). The endogenous protein should appear around the anticipated 213 kDa.

Bioinformatics

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