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Our Abpromise guarantee covers the use of ab22651 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).|
|ELISA||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use at an assay dependent concentration. PubMed: 20436478|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 20436478|
|Flow Cyt||Use 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
ab22651 staining Vimentin in Ferret brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with Triton-X and blocked with 7.5ug/ml 0.1 M glycine buffer for 30 minutes at room temperature. Samples were incubated with primary antibody (1/50) for 16 hours at 4°C. A Cy2®-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
Western Blot analysis using ab22651
ab22651 at 1µg/ml staining Vimentin (phospho S55) in human U251 cells by Immunocytochemistry/ Immunofluorescence.
Overlay histogram showing HeLa cells stained with ab22651 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab22651, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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