Anti-Acetylated alpha Tubulin antibody [6-11B-1] (ab24610)

  Western blot - Acetylated alpha Tubulin antibody [6-11B-1] (ab24610)

All lanes : Anti-Acetylated alpha Tubulin antibody [6-11B-1] (ab24610) at 5 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : Brain (Mouse) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size : 55 kDa
Observed band size : 55 kDa
Additional bands at : 140 kDa,25 kDa,35 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 150 seconds

  Immunocytochemistry/ Immunofluorescence - Acetylated alpha Tubulin antibody [6-11B-1] (ab24610)

ICC/IF image of ab24610 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab24610, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Acetylated alpha Tubulin antibody [6-11B-1] (ab24610)

ab24610 at 1/100 dilution staining acetylated alpha tubulinin in prostate carcinoma by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were paraformaldehyde fixed, permeabilized in Triton X-100 prior to blocking in 1% serum for 1 hour at 27°C and then incubated with ab24610 for 12 hours at 4°C. Alexa fluor® 546 donkey polyclonal to mouse Ig, diluted 1/500, was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

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  Immunocytochemistry/ Immunofluorescence - Acetylated alpha Tubulin antibody [6-11B-1] (ab24610)

ICC/IF image of ab24610 stained HepG2 cells. The cells were 4% PFA 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 (ab24610, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.

  Flow Cytometry-Anti-Acetylated alpha Tubulin antibody [6-11B-1](ab24610)

Overlay histogram showing HeLa cells stained with ab24610 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab24610, 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 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.