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Our Abpromise guarantee covers the use of ab24525 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 22919071|
|WB||Use at an assay dependent concentration.|
Immunohistochemical analysis of murine eye tissue, staining Vimentin (red) with ab24525.
Tissue was fixed in paraformaldehyde, blocked for 1 h at room temperature in 0.5% BSA, 0.2% Tween-20 plus 5% goat serum. Samples were incubated with primary antibody (1/200) for 1 hour at room temperature. A Cy3-conjugated goat anti-chicken IgG (1/200) was used as the secondary antibody.
Rat cerebral cortex cultures stained with chicken antibody to vimentin ab24525 (green) and rabbit antibody to GFAP (red). Note flattened fibroblastic cells are mostly green (i.e. vimentin positive, GFAP negative), while clearly astrocytic cells, express both vimentin and GFAP and therefore appear golden or orange. Certain other cells express predominantly GFAP and therefore appear red.
ab24525 staining Vimentin in rat smooth muscle cells from mesenteric artery by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 4% paraformaldehyde in physiological saline solution (PSS) 4 min at 4°C and permeabilized with 0.3% Triton x100 before blocking with 2% BSA was done for 30 minutes at 20°C. Samples were incubated with primary antibody (1/300: in PSS with 2%BSA and 0.3% Triton X-100) for 14 hours at 4°C. An Abcam`s ab6875, goat anti-chicken IgY Texas Red was used as secondary antibody at 1/400 dilution.
Western blot of Rat whole brain extract,HeLa,SH-SY5Y,HEK293, and NIH/3T3 cells probed with ab24525,showing a single strong band at ~ 50kDa.