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Rabbits were immunized with a phosphorylated synthetic peptide, which represented the portion of human Structural Maintenance of Chromosomes-1 (LocusLink ID 8243)around Serine 966, conjugated to KLH. After multiple immunizations in Freund's adjuvant, serum was collected.
Our Abpromise guarantee covers the use of ab1276 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|IP||Use at 2-6 µg/mg of lysate.|
|WB||1/500 - 1/5000. Detects a band of approximately 160 kDa (predicted molecular weight: 143 kDa).|
|ICC/IF||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
IHC image of SMC1 (phospho S966) staining in human spleen 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 ab1276, 5µg/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.