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Our Abpromise guarantee covers the use of ab78316 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IP||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 53 kDa.|
|ELISA||Use at an assay dependent concentration.|
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 UHRF1, which results in an increase in acetylation of p53 at Lys120. Total p53 was immunoprecipitated with anti-p53 monoclonal antibody from the crude extracts and analyzed by western blotting with another anti-p53 antibody (upper panel) and ab78316, at 1µg/ml (middle panel). The lower panel shows total p53.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with 100 nM Doxorubicin for 24 hour labelling p53 (acetyl K120) with ab78316 at a dilution of 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.25% Triton X-100 in PBS for 10 minutes. DAPI was used to stain the nucleus.
Crude whole cell extracts were prepared from H129 cells (p53 negative cell line) expressing only Myc-p53 (first lane), and both Myc-p53 and His-Tip60 (Second lane). In the upper panel, the cell extracts were immuno-blotted with anti-Myc, anti-His-tag or anti-α-tublin antibodies. In the lower panel, the extracts were immuno-precipitated with ab78316 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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