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Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause.
Our Abpromise guarantee covers the use of ab684 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. PubMed: 17478566Use at an assay dependent dilution.|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
Blocked with 5% milk for 1 hour at 20°C.
Incubated with the primary antibody in 5% milk for 16 hours at 4°C.
The 4 lanes were each loaded with 10ug of protein based upon a BCA assay for protein quantification. However, looking at a subsequent loading control blot (vinvulin) lanes 1 and 4 are under loaded versus lanes 2 and 3.
ab684 staining non-muscle Myosin IIB in WIF-B cells (hepatoma-derived hybrid cell line) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1% donkey serum in 1% PBST for 1 hour at 22°C. Samples were incubated with primary antibody (1/50 in blocking buffer) for 3 hours at 22°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.
Overlay histogram showing HeLa cells stained with ab684 (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. The cells were then incubated with the antibody (ab684, 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 in 0.1% PBS-Triton used under the same conditions.