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53BP1 [p Ser25] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NB100-1803

Novus Biologicals, part of Bio-Techne
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NB100-1803

Key Product Details

Validated by

Independent Antibodies, Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Cited:

Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

0.2 mg/ml

Product Specifications

Immunogen

53BP1 [p Ser25] Antibody was made to a phosphorylated synthetic peptide, which represented a portion of human p53 Binding Protein 1 (GeneID 7158) around serine 25 according to the numbering given in entry NP_005648.1.

Modification

p Ser25

Marker

DNA Double Strand Break Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for 53BP1 [p Ser25] Antibody

Western Blot: 53BP1 [p Ser25] Antibody [NB100-1803]

Western Blot: 53BP1 [p Ser25] Antibody [NB100-1803]

Western Blot: 53BP1 [p Ser25] Antibody [NB100-1803] - Samples: Whole cell lysate (30 ug or 15 ug for WB; 1 mg for IP, 20% of IP loaded) from 293T cells that were mock treated or treated with NCS (200 ng/ml; 1h). Affinity purified rabbit anti-Phospho 53BP1 antibody at 0.04 ug/ml (A) and 0.1 ug/ml (B) for WB. 53BP1 was immunoprecipitated using affinity purified rabbit anti-53BP1 Antibody (Catalog #NB100-304). Detection: Chemiluminescence with exposure times of 3 seconds (A and B). Bands appear at ~350 kDa as the observed molecular weight and the theoretical molecular weight is 214 kDa.
Flow Cytometry: 53BP1 [p Ser25] Antibody [NB100-1803]

Flow Cytometry: 53BP1 [p Ser25] Antibody [NB100-1803]

Flow Cytometry: 53BP1 [p Ser25] Antibody [NB100-1803] - Flow cytometric analysis of phospho 53BP1 (S25). Jurkat cells were treated with neocarzinostatin (NCS) for 1.5 hrs. 1 Million cells were fixed, permeabilized, and stained with 1.5 ug/ml antiphospho 53BP1 (S25) (Catalog #NB100-1803) in a 150 ul reaction. Isotype control (black), untreated (red), NCS treated (blue).
Immunohistochemistry-Paraffin: 53BP1 [p Ser25] Antibody [NB100-1803]

Immunohistochemistry-Paraffin: 53BP1 [p Ser25] Antibody [NB100-1803]

Immunohistochemistry-Paraffin: 53BP1 [p Ser25] Antibody [NB100-1803] - FFPE sections of human stomach carcinoma (left) and mouse teratoma (right). Antibody: Affinity purified rabbit anti- Phospho 53BP1 (S25) used at a dilution of 1:1,000 (1ug/ml). Detection: DAB.

Applications for 53BP1 [p Ser25] Antibody

Application
Recommended Usage

Flow Cytometry

25 ug per 1 million cells in a 150 mcl reaction

Immunocytochemistry/Immunofluorescence

1:500 - 1:2500

Immunohistochemistry

1:1000 - 1:5000

Immunohistochemistry-Paraffin

1:1000 - 1:5000

Immunoprecipitation

1-4 ug/mg of lysate

Western Blot

1:2000-1:10000
Application Notes
Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections. ICC/IF reactivity reported in (PMID: 27713178).

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

TBS and 0.1% BSA

Preservative

0.09% Sodium Azide

Concentration

0.2 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. Do not freeze.

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

Entrez Gene IDs

7158 (Human)

Gene Symbol

TP53BP1

UniProt

Additional 53BP1 Products

Product Documents for 53BP1 [p Ser25] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for 53BP1 [p Ser25] Antibody

This antibody can be used as the primary antibody in a PLA assay with the following as complementing antibodies: NB100-97831, NB110-40543, NB100-322, NB200-171, NB100-68265

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.

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