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Tissue/ cell preparation (Human). (HPB-ALL T cell line)
Our Abpromise guarantee covers the use of ab8219 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use a concentration of 2 µg/ml.
Use at an assay dependant concentration. For detection of activated platelets, neutrophils and basophils.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use a concentration of 10 µg/ml.|
|ICC/IF||Use at an assay dependent concentration.|
ab8219 staining CD63 in human fibrosarcoma cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde. Samples were incubated with primary antibody which was used diluted to 10 µg/ml for 1 hour at 20°C. An Alexa Fluor®555-conjugated donkey monoclonal to mouse IgG was used at dilution at 1/500 as secondary antibody.
IHC image of CD63 staining in Human skin melanoma 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 ab8219, 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.