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Our Abpromise guarantee covers the use of ab9964 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 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/500. Detects a band of approximately 28, 40 kDa (predicted molecular weight: 28 kDa). (see Erdely et al and recommended protocol). Block membrane in 5% milk and incubate antibody overnight in TBST containing 1% milk. Erdely HA et al noted that "...a much lighter intensity band was also detected at approximately 40 kDa, which unlike the band at 28 kDa, was not blocked by the blocking peptide. Therefore, the 40 kDa band was considered to be nonspecific."|
IHC image of ab9964 staining in human normal cerebral cortex 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 ab9964, 1/1000 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.