The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/10000. Detects a band of approximately >220 kDa (predicted molecular weight: 214 kDa).
Use at 1-4 µg/mg of lysate.
Use 0.25µg for 106 cells.
0.25 μg per 1 million cells in a 150 mcl reaction.
ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
Post-translational 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma (left) and mouse teratoma (right) tissues labelling 53BP1 (phospho S24) with ab70323 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - 53BP1 (phospho S25) antibody (ab70323)
All lanes : Anti-53BP1 (phospho S25) antibody (ab70323) at 0.1 µg/ml
Lane 1 : Whole cell lysate from 293T cells, untreated Lane 2 : Whole cell lysate from 293T cells treated
with NCS (200 ng/ml for 1h)