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Fusion protein (Human).
Our Abpromise guarantee covers the use of ab28 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.|
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).|
|IP||Use at 10 µg/mg of lysate.|
|ELISA||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|RIA||Use at an assay dependent concentration.|
|IHC-Fr||1/100 - 1/250.|
IHC image of p53 staining in human pancreas 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 ab28, 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.
This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab28 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
The image shows a Western blot for ab28, testing the threshold detection of the p53 antibody. The lanes are as follows:
Lane 1 - 20ug of A549 lysate, lane 2 - 10ug of A549 lysate, lane 3 - 5ug of A549 lysate, lane 4 - 2.5ug of A549 lysate, lane 5 - 1.5ug of A549 lysate.
This picture was submitted as part of the review by Craig Carson.
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