53BP1 Antibody (1285C) was made to a synthetic peptide made to the C-terminal portion of human 53BP1 protein (between amino acids 1900-1972) [UniProt Q12888]

Reactivity Notes

Predicted cross-reactivity based on sequence identity: Bovine (100%), Canine (100%), Chimpanzee (100%), Equine (100%), Feline (100%), Gibbon (100%), Gorilla (100%), Hamster (100%), Marmoset (100%), Orangutan (100%), Panda (100%), Porcine (100%), Rabbit (100%), Sheep (100%), Bat (99%), Elephant (99%), Rat (99%)

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for 53BP1 Antibody (1285C) - BSA Free

Immunohistochemistry-Paraffin: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunohistochemistry-Paraffin: 53BP1 Antibody (1285C) [NBP2-54753] - 53BP1 was detected in paraffin-embedded sections of human cervical cancer tissue using Rabbit Anti-Human 53BP1 Monoclonal Antibody (clone 1285C) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei.

Western Blot: 53BP1 Antibody (1285C)BSA Free [NBP2-54753]

Western Blot: 53BP1 Antibody (1285C) [NBP2-54753] - Total protein from human A431, NTERA-2, HeLa and mouse 3T3 cell lines was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/ml 53BP1 Antibody in block solution and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. The observed molecular weight is shown on this gel at ~250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - Neuro2a cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #NB100-690) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow (Intracellular): 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Flow (Intracellular): 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on HeLa Cells with 53BP1 (1285C) antibody (Catalog #NBP2-54753) (blue) and a matched isotype control MAB1050 (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody [1285C] NBP2-54753 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Flow Cytometry: 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody [1285C] NBP2-54753 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #NB100-690) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - NIH-3T3 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #NB100-690) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody [1285C] NBP2-54753 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow (Intracellular): 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Flow (Intracellular): 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on HeLa cells with 53BP1 [1285C] Antibody (Catalog #NBP2-54753AF647) (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. Image using the Alexa Fluor 647 format of this antibody.

53BP1 (1285C) in U-2 OS Human Cell Line.

53BP1 (1285C) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-53BP1 (1285C) Protein G Purified Recombinant Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NBP2-54753AF488) (green) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

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Applications for 53BP1 Antibody (1285C) - BSA Free

Application

Recommended Usage

Flow (Intracellular)

1 ug/mL

Flow Cytometry

2.5-5 ug/ml

Immunohistochemistry

1.0 ug/ml

Immunohistochemistry-Paraffin

1.0 ug/ml

Western Blot

1.0 ug/ml

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: 53BP1

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

Long Name

p53 Binding Protein 1

Alternate Names

p202, TP53BP1

Gene Symbol

TP53BP1

Additional 53BP1 Products

Product Documents for 53BP1 Antibody (1285C) - BSA Free

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for 53BP1 Antibody (1285C) - BSA Free

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.

Citations
Reviews

53BP1 Antibody (1285C) - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-54753

Key Product Details

Species Reactivity

Validated:

Cited:

Predicted:

Applications

Validated:

Cited:

Label

Antibody Source

Format

Concentration

View all 53BP1 Antibodies »

Product Specifications

Immunogen

Reactivity Notes

Clonality

Host

Isotype

Scientific Data Images for 53BP1 Antibody (1285C) - BSA Free

Immunohistochemistry-Paraffin: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Western Blot: 53BP1 Antibody (1285C)BSA Free [NBP2-54753]

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Flow (Intracellular): 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Flow Cytometry: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

Flow (Intracellular): 53BP1 Antibody (1285C) - BSA Free [NBP2-54753]

53BP1 (1285C) in U-2 OS Human Cell Line.

Applications for 53BP1 Antibody (1285C) - BSA Free

Flow (Intracellular)

Flow Cytometry

Immunohistochemistry

Immunohistochemistry-Paraffin

Western Blot

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: 53BP1

Long Name

Alternate Names

Gene Symbol

Additional 53BP1 Products

Product Documents for 53BP1 Antibody (1285C) - BSA Free

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for 53BP1 Antibody (1285C) - BSA Free