Anti-p53 Ac-K120 (human) antibody, mouse monoclonal (10E5)
Product code71-131

Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium.

ImmunogenSynthetic peptide containing acetyl-Lys315 of human p53

Isotype

Mouse IgG1κ

Reactivity

Human.

Special notes

N/A

Application

1. Western blotting (~1 µg/ml) (Fig.1)
2. Immuno-precipitation (Fig.2)
3. Immunofluorescence staining (Fig.3)
4. Flow-Cytometry (1/100)
5. ELISA

Backgroundp53 mutants are found in more than half of human cancers and are consideredas the most important human cancer related gene. p53 is detected at 53kD
position by electrophoresis and is composed of 393 amino acids. In theunstressed normal cells, the p53 level is low and it is inactive. However, withstress, especially with DNA damage, it is activated to promote arrest of cell cycle
and repair of DNA damage, or induction of apoptosis. The functions and stabilityof p53 are regulated by phosphorylation of serine and threonine, and acetylationof lysine at various sites in the molecule.
Acetylation of lysine 120 (acetyl-K120) of p53 occurs rapidly after DNA
damage and is catalyzed by the MYST family acetyltransferases hMOF andTIP60, and activates transcription of proapoptotic genes, BAX and PUMA.

Data Link

UniProtKB/Swiss-Prot P04637 (P53_HUMAN)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 71-131 Anti-p53 Ac-K120 (human) antibody, mouse monoclonal (10E5)

Fig.１ Identification of p53 protein, whose Lys 120 is acetylated,by Western blotting with 10E5

monoclonal antibody.
Samples are crude lysates of HCT116 cells: Left lanes are control. Right lanes are cells treated withsiRNA to knockdown the expression of a Tip60 interacting protein UHRF1, which results in increasein acetylation of p53 at Lys120. Total p53 was immuno-precipitated with omnipotent anti-p53
monoclonal antibody (DO-1) from the crude extracts and analyzed by Western blotting with anti-p53
antibody (FL393) (upper panel) or anti-p53 acetyl-K120 monoclonal antibody (10E5) (middle panel).
The lower panel shows total p53.

Fig.2 Immunoprecipitation of p53 acetylated at K120 by 10E5 monoclonal antibody.
Crude cell extracts were prepared from H129 cells (p53 negative cell line) expressing only Myc-p53(first lane), and both Myc-p53 and His-Tip60. In the upper panel, the whole cell extracts wereimmuno-blotted with anti-Myc, anti-His-tag or anti-α-tublin antibodies. In the lower panel, theextracts were immuno-precipitated with anti-p53 Ac-K120 antibody (10E5) and the precipitates wereimmuno-blotted with anti-Myc antibody. Acetylation of p53 at K120 is dependent on
Tip60 and promoted by over-expression of His-Tip60.
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Fig.3. Immunofluorescence staining of p53 acetyl-K120 in nuclei of HeLa cells subjected to DNAdamage.
HeLa cells were treated with 100 nM Doxorubicin for 24 hr, fixed with 4% paraformaldehyde
overnight, permeabilized with 0.25% Triton X-100 in PBS for 10 min.
The antibody was used at 1/1,000 dilution
Nucleus (DNA) was stained with DAPI

References : This antibody has been used in the following publication.
1. Rokudai S et al. MOZ increases p53 acetylation and premature senescence through its complex
formation with PML.Proc Natl Acad Sci U S A. 2013 Mar 5;110(10):3895-900. PMID: 23431171.
WB (human)
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BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/