53BP1 Recombinant Rabbit Monoclonal Antibody [JA66-11]
Overview
Product Name
53BP1 Recombinant Rabbit Monoclonal Antibody [JA66-11]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human 53BP1 aa 1-50 / 1972.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC
Target Molecular Weight
Predicted band size: 214 kDa
Positive Control
HeLa cell lysate, HT-29 cell lysate, human brain tissue lysate, mouse brain tissue lysate, rat heart tissue lysate, HeLa, human brain tissue, mouse spleen tissue, mouse testis tissue, rat testis tissue, human spleen tissue, human liver tissue, human cervix tissue.
Conjugation
unconjugated
Clone Number
JA66-11
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:2,000
-
IF-Cell
-
1:50
-
IF-Tissue
-
1:200
-
IHC-P
-
1:200-1:5,000
-
FC
-
1:1,000
Target
Function
The p53 binding proteins 53BP1 and 53BP2 (Bbp) bind to the central DNA-binding domain of wild type p53, but do not bind mutant p53. The central DNA-binding domain of p53 is required for site-specific DNA binding and is frequently mutated in malignant tumors. Binding of 53BP1 to the L3 loop of p53 and of 53BP2 to the L2 loop of p53 confirms that the loop is dependent on p53 conformation. Site-specific binding also suggests that 53BP1 and 53BP2 are involved in p53-mediated tumor suppression. 53BP1 was isolated from H258 cells and is expressed in Jurkat cells in both the cytoplasm and the nucleus. The N-terminus of 53BP2 is localized to the cytoplasm, while the C-terminus might be localized in the nucleus. 53BP1 promotes cell proliferation by binding to p202, whereas 53BP2 induces cell death by binding to Bcl2 and NFkB p65.
Background References
1. Manchon JF et al. Levetiracetam mitigates doxorubicin-induced DNA and synaptic damage in neurons. Sci Rep 6:25705 (2016).
2. Alessio N et al. Low dose radiation induced senescence of human mesenchymal stromal cells and impaired the autophagy process. Oncotarget 6:8155-66 (2015).
Post-translational Modification
Asymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.; Phosphorylated at basal level in the absence of DNA damage. Phosphorylated by ATM in response to DNA damage: phosphorylation at different sites promotes interaction with different set of proteins: phosphorylation at the N-terminus by ATM (residues from 6-178) promotes interaction with PAXIP1 and non-homologous end joining (NHEJ) of dysfunctional telomeres. Phosphorylation by ATM at residues that are located more C-terminus (residues 300-650) leads to promote interaction with RIF1. Interaction with RIF1 leads to disrupt interaction with NUDT16L1/TIRR. Phosphorylation at Thr-1609 and Ser-1618 in the UDR motif blocks interaction with H2AK15ub. Dephosphorylated by PPP4C. Hyperphosphorylation during mitosis correlates with its exclusion from chromatin and DNA lesions. Hyperphosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation. Dephosphorylated by PPP5C.
Subcellular Location
Nucleus. Chromosome.
Synonyms
53 BP1 antibody
53BP1 antibody
FLJ41424 antibody
MGC138366 antibody
p202 antibody
p53 binding protein 1 antibody
p53 BP1 antibody
p53-binding protein 1 antibody
p53BP1 antibody
TP53 BP1 antibody
ExpandImages
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Western blot analysis of 53BP1 on different lysates with Rabbit anti-53BP1 antibody (ET1704-05) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HT-29 cell lysate (15 µg/Lane)
Lane 3: Human brain tissue lysate (20 µg/Lane)
Lane 4: Mouse brain tissue lysate (20 µg/Lane)
Lane 5: Rat heart tissue lysate (20 µg/Lane)
Predicted band size: 214 kDa
Observed band size: 450 kDa
Exposure time: 43 seconds; ECL: K1801;
3-8% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-05) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling 53BP1 with Rabbit anti-53BP1 antibody (ET1704-05) at 1/50 dilution.
Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-53BP1 antibody (ET1704-05) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human cervix tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunofluorescence analysis of paraffin-embedded human brain tissue labeling 53BP1 with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-05, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). -
Immunofluorescence analysis of paraffin-embedded mouse testis tissue labeling 53BP1 with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-05, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). -
Flow cytometric analysis of HeLa cells labeling 53BP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1704-05, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
-
Preclinical Study of CDH3-Targeted 89Zr/177Lu Theranostics in Triple-Negative Breast Cancer
Journal: Molecular Pharmaceutics
DOI: 10.1021/acs.molpharmaceut.5c01584
IF: 4.5
Application: IF-tissue
Reactivity: Mouse
Publish date: 2026 Jan
-
tRNA-derived fragment tRF-24 drives CELF1 phase separation to promote oncogenic splicing in esophageal squamous cell carcinoma
Journal: Journal Of Experimental & Clinical Cancer Research
DOI: 10.1186/s13046-025-03553-x
IF: 12.8
Application: IF-cell
Reactivity: Human
Publish date: 2025 Nov
-
Discovery of Bi-magnolignan as a novel BRD4 inhibitor inducing apoptosis and DNA damage for cancer therapy
Journal: Biochemical Pharmacology
DOI: 10.1016/j.bcp.2025.116843
IF: 5.3
Application:
Reactivity:
Publish date: 2025 Feb
-
ARIH1 activates STING-mediated T-cell activation and sensitizes tumors to immune checkpoint blockade
Journal: Nature Communications
DOI:
IF: 16.6
Application: IF-cell
Reactivity: Human
Publish date: 2023 Jul
-
A novel and potent dihydroorotate dehydrogenase inhibitor suppresses the proliferation of colorectal cancer by inducing mitochondrial dysfunction and DNA damage
Journal: Medcomm-Oncology
DOI:
IF: NA
Application: WB
Reactivity: Human
Publish date: 2022 Jul
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