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A cytoskeletal preparation of human callus material was used for immunization.
Our Abpromise guarantee covers the use of ab18586 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 2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|ICC||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 60 kDa.|
|IHC-P||1/10. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.|
|IHC-Fr||Use at an assay dependent concentration.|
ab18586 staining Cytokeratin 6 in human breast gland tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with methanol and blocked with 10% serum for 5 minutes. The sample was incubated with primary antibody (1/25) for 1 hour. An HRP-conjugated rabbit polyclonal to mouse IgG was used undiluted as secondary antibody. Ultravision ONE detection system has been used.
Overlay histogram showing A431 cells stained with ab18586 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18586, 2µg/1x106 cells ) for 30 min at 22°C. The secondary antibody used was a goat anti-mous DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.
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