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Synthetic peptides, corresponding to amino acids 20-34 (GWKQGDEEEKWAEKA) and 316-330 (VNRNSTIENTRRHIG) of Human SMAD 5. These sequences are conserved in human Smad 1, 8 and 9 and expected to cross react due to sequence homology.
Our Abpromise guarantee covers the use of ab13724 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/2000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).|
|IP||Use at an assay dependent concentration.|
Image from Tao Shijie et. al., Development 136, 2757-2765 (2009), Fig S4.
IHC image of SMAD5 staining in Human normal lung 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 ab13724, 1/200 dilution, 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.