Anti p53, acetyl Lys120 - Cosmo Bio Co.,Ltd.



Anti p53, acetyl Lys120 Antibody

Catalog No.: BAM-71-131-EX
Size: 50UG
[1 mg / mL]
Price: see also
Size 100
antigen/source: p53
host: Mouse
Label: Unlabeled
Purity: IgG fraction
Application: Western Blot
Enzyme Linked Immunosorbent Assay
catalog info: Catalog 2012-p73
Storage: -20C
Immunogen: Human
Isotype: IgG1
Clone: 10E5
Reacts with: Human


Fig.1 Identification of p53 protein, whose Lys120 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 with siRNA to knockdown the expression of a Tip60 interacting protein, which results in increase in 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 were immuno-blotted with anti-Myc, anti-His-tag or anti-αtublin antibodies. In the lower panel, the extracts were immuno-precipitated with anti-p53 Ac-K120 antibody (10E5) and the precipitates were immuno-blotted with anti-Myc antibody. Acetylation of p53 at K120 is dependent on Tip60 and promoted by over-expression of His-Tip60.


Fig.3. Immunofluorescence staining of p53 acetyl-K120 in nuclei of HeLa cells subjected to DNA damage. 1. HeLa cells were treated with 100 nM Doxorubicin for 24 hr, fixed with 4% paraformaldehyde overnight, permealized with 0.25% Triton X-100 in PBS for 10 min. 2. Incubate cells with 1.5% BSA in PBS for 30 min to block unspecific binding of the antibodies. Incubate the cells with 1/1,000 diluted anti-p53(acetyl-K120) antibody in 1% BSA in PBS at 4°C overnight. 3. Incubate cells with the secondary antibody, goat anti-mouse IgG conjugated with Alex 488, at 1/1,000 dilution in 1% BSA for 1 hr at room temperature. 4. Nucleus (DNA) was stained with DAPI