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Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human N Cadherin.
(Peptide available as ab18620.)
Our Abpromise guarantee covers the use of ab18203 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 125-135 kDa (predicted molecular weight: 100 kDa).|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use a concentration of 10 µg/ml.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
This blot was produced using a 4-12% 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 2% Bovine Serum Albumin before being incubated with ab18203 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
The N Cadherin protein has a predicted molecular weight of 100 kDa, however it is extensively glycosylated and has been shown to run in the 125-135 kDa region (SwissProt data).
IHC image of N Cadherin staining in Human liver cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab18203, 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.