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Native chick brain microtubules.
Our Abpromise guarantee covers the use of ab80779 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 1µg for 106 cells.|
|WB||Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 54 kDa.|
|IP||Use at 2 µg/mg of lysate.|
|Electron Microscopy||Use at an assay dependent dilution.|
|ICC/IF||Use at an assay dependent dilution.|
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab80779 overnight at 4°C. Antibody binding was detected using Anti-rabbit Alexa Fluor® 790 (ab175781) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
ab80779 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab80779 at 5µg/ml overnight at +4°C. The secondary antibody (green) was anti-mouse DyLight® 488 (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in methanol fixed (100%, 5min) HeLa, Hek293, HepG2, and MCF-7 cells, also in formaldehyde fixed (4%, 10min) HeLa, Hek293, and MCF-7 cells at 5ug/ml.
Formalin-fixed, paraffin-embedded Human lung stained with ab80779 at 1/400 dilution using a peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of ciliated epithelium of bronchioles.
Overlay histogram showing HEK293 cells stained with ab80779 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab80779, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was anti-mouse DyLight® 488 (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 HEK293 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.
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