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Synthetic peptide corresponding to Human MDM2 aa 154-167. Cross-linked to KLH using glutaldehyde.
Our Abpromise guarantee covers the use of ab3110 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
IHC image of MDM2 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3110, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab3110 staining MDM2 in human pancreas tissue (ab29816) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval in citric buffer pH 6.0, prior to blocking with 10% serum for 1 hour at 12°C. The primary antibody was diluted 1/100 and incubated with the sample for 12 hours at 4°C. An HRP-conjugated rabbit anti-mouse polyclonal was used as the secondary antibody, diluted 1/200.
Overlay histogram showing HeLa cells stained with ab3110 (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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3110, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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