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A trial size is available to purchase for this antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Our Abpromise guarantee covers the use of ab62376 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
For unpurified use at 1/50 000 - 1/100 000.
|IHC-P||1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
For unpurified use at 1/100 - 1/250.
|ICC/IF||1/100 - 1/250.|
For unpurified use at 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Immunocytochemistry/Immunofluorescence analysis of HepG2 +/- (etoposide + trichostatin A) (etoposide 30ug/ml 8hr followed by TSA 500ng/ml 4hr) cells labelling p53 (acetyl K373) with purified ab62376 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291 anti-Tubulin (mouse mAb) primary and ab150120 (AlexaFluor®594 goat anti-mouse) secondary both at 1/1000 dilution. Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody and ab150120 (anti-mouse) secondary antibody were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by ab150077 (anti-rabbit secondary antibody).
Immunohhistochemical analysis of Paraffin-embedded human breast ductal carcinoma tissue sections labelling p53 with unpurified ab62376 at 1/100 dilution.
Immunohistochemical analysis of paraffin-embedded human cervical carcinoma sections labelling p53 (acetyl K373) with purified ab62376 at dilution of 1/800. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Flow cytometry analysis of HeLa cells labelling p53 (acetyl K373) (red) with purified ab62376 at dilution of 1/120. The secondary antibody used was goat anti rabbit IgG (FITC) at 1/150. Cells were fixed with 4% paraformaldehyde. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.
Overlay histogram showing HeLa cells stained with unpurified ab62376 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab62376, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG; H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.