Product nameAnti-53BP1 antibody
See all 53BP1 primary antibodies
DescriptionRabbit polyclonal to 53BP1
Tested applicationsSuitable for: IP, IHC-P, WB, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat, Rabbit, Cow, Baboon
Synthetic peptide corresponding to Human 53BP1 aa 1950 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; HepG2; HEK293.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab87097 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 213 kDa (predicted molecular weight: 213 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
FunctionMay have a role in checkpoint signaling during mitosis. Enhances TP53-mediated transcriptional activation. Plays a role in the response to DNA damage.
Involvement in diseaseNote=A chromosomal aberration involving TP53BP1 is found in a form of myeloproliferative disorder chronic with eosinophilia. Translocation t(5;15)(q33;q22) with PDGFRB creating a TP53BP1-PDGFRB fusion protein.
Sequence similaritiesContains 2 BRCT domains.
modificationsAsymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.
Phosphorylated at basal level in the absence of DNA damage. Hyper-phosphorylated in an ATM-dependent manner in response to DNA damage induced by ionizing radiation. Hyper-phosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation.
Cellular localizationNucleus. Chromosome > centromere > kinetochore. Associated with kinetochores. Both nuclear and cytoplasmic in some cells. Recruited to sites of DNA damage, such as double stand breaks. Methylation of histone H4 at 'Lys-20' is required for efficient localization to double strand breaks.
- Information by UniProt
- 53 BP1 antibody
- 53BP1 antibody
- FLJ41424 antibody
ICC/IF image of ab87097 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab87097, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-53BP1 antibody (ab87097) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 213 kDa
Observed band size: 213 kDa
Exposure time: 5 minutes
53BP1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to 53BP1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab87097.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 213kDa: 53BP1.
IHC image of 53BP1 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab87097, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
This product has been referenced in:
- Cadoni E et al. Caloric restriction delays early phases of carcinogenesis via effects on the tissue microenvironment. Oncotarget 8:36020-36032 (2017). WB . Read more (PubMed: 28415598) »
- Carvalho S et al. SETD2 is required for DNA double-strand break repair and activation of the p53-mediated checkpoint. Elife 3:e02482 (2014). IP ; Human . Read more (PubMed: 24843002) »