Cells were grown in liquid TAP media 18 hours prior to the experiment. Cells were incubated with either 10 μM LatB or 1% DMSO for 10, 30, and 120 min. Cells were fixed to coverslips with methanol fixation and washed once with 1X PBS. 100% block was added to the coverslips for 30 min and then replaced with block with 10% normal goat serum for an additional 30 mins. The primary antibodies, rat anti-HA antibody (Sigma) at 1:1000 dilution and rabbit anti-acetylated alpha tubulin at 1:1000 dilution antibody (Cell Signaling Technology) were added to the coverslips and placed in the 4°C overnight. The primary antibody was removed and 4 X 5 min wash in 1 X PBS. The anti-rat and anti-rabbit secondary antibodies (Sigma) were added to the coverslips at 1:1000 dilution for 1 hour. The secondary antibodies were removed and another 4 X 5 min wash in 1 X PBS. Coverslips were mounted with Fluoro-mount-G. Slides were visualized on Nikon TiS microscope on the FITC, TRITC, and brightfield channels.
Rabbit anti acetylated alpha tubulin
Manufactured by Cell Signaling Technology
Rabbit anti-acetylated alpha tubulin is a primary antibody that specifically binds to the acetylated form of the alpha subunit of tubulin, a key component of the cytoskeleton. It can be used to detect and quantify the level of acetylated alpha tubulin in various cell and tissue samples.
Automatically generated - may contain errors
2 protocols using rabbit anti acetylated alpha tubulin
1
Latrunculin B Actin Disruption Assay
2
Latrunculin B Actin Disruption Assay
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Cells were grown in liquid TAP media 18 hours prior to the experiment. Cells were incubated with either 10 μM LatB or 1% DMSO for 10, 30, and 120 min. Cells were fixed to coverslips with methanol fixation and washed once with 1X PBS. 100% block was added to the coverslips for 30 min and then replaced with block with 10% normal goat serum for an additional 30 mins. The primary antibodies, rat anti-HA antibody (Sigma) at 1:1000 dilution and rabbit anti-acetylated alpha tubulin at 1:1000 dilution antibody (Cell Signaling Technology) were added to the coverslips and placed in the 4°C overnight. The primary antibody was removed and 4 X 5 min wash in 1 X PBS. The anti-rat and anti-rabbit secondary antibodies (Sigma) were added to the coverslips at 1:1000 dilution for 1 hour. The secondary antibodies were removed and another 4 X 5 min wash in 1 X PBS. Coverslips were mounted with Fluoro-mount-G. Slides were visualized on Nikon TiS microscope on the FITC, TRITC, and brightfield channels.
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