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

Antibodies

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Anti p53, acetyl Lys120 Antibody

Catalog No.: BAM-71-131-EX
Size: 50UG
[1 mg / mL]
Price: see also
#BAM-71131EX
Size 100
antigen/source: p53
host: Mouse
Label: Unlabeled
Purity: IgG fraction
Application: Western Blot
Enzyme Linked Immunosorbent Assay
Immunoprecipitation
catalog info: Catalog 2012-p73
Storage: -20C
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Immunogen: Human
Isotype: IgG1
Clone: 10E5
Reacts with: Human



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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.

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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.

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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
 
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