Mouse anti α tubulin antibody

Manufactured by Thermo Fisher Scientific

Sourced in United States

The mouse anti-α-tubulin antibody is a laboratory reagent used to detect and visualize α-tubulin, a component of the cytoskeleton in eukaryotic cells. It can be used in various cell biology techniques, such as Western blotting, immunocytochemistry, and immunohistochemistry, to study the distribution and dynamics of the cytoskeleton.

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Lab products found in correlation

Lsm 700, by Zeiss (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Methanol acetone, by Merck Group (1 mentions) Tcs sp8 confocal laser scanning microscope, by Leica (1 mentions) Alexa fluor 594 goat anti mouse igg h l antibody, by Thermo Fisher Scientific (1 mentions) Formalin neutral buffer solution, by Merck Group (1 mentions) Nis element, by Nikon (1 mentions)

3 protocols using mouse anti α tubulin antibody

Immunofluorescence Analysis of Parasite Cytoskeleton

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Parasites treated with DMSO (the negative control) or terfenadine (1, 2, 3 μM) were harvested after 48 h by centrifugation (10 min at 1973× g at 4 °C), washed twice in PBS, and allowed to attach to polyethylenimine-coated coverslips for 20 min. The coverslips were fixed with methanol–acetone (Sigma-Aldrich, St. Louis, MO, USA) in a 1:1 ratio at −20 °C for 10 min. The adhered cells were also permeabilized with 0.05% Triton X-100 (in PBS) for 30 min. After two washes with PBS, cells were incubated for 1 h with 1% bovine serum albumin (BSA) to block nonspecific binding. Next, cells were incubated with diluted 1:1000 mouse anti-α-tubulin antibody (Invitrogen, Thermo Fisher, Scientific, Waltham, MA, USA) for 1 h at room temperature. After two PBS washes, the cells were incubated with goat anti-mouse IgG and FITC conjugated (1:200 dilution, Thermo Fisher Scientific). The coverslips were washed 10 times in PBS and then mounted on microscope slides with a drop of mounting medium containing DAPI (Prolong Gold Invitrogen). The cells were analyzed using a Leica confocal microscope (Leica TCS SP8, Confocal Laser Scanning Microscope), and images were processed using Leica Lite Software.

Suárez-Rico D.O., Munguía-Huizar F.J., Cortés-Zárate R., Hernández-Hernández J.M., González-Pozos S., Perez-Rangel A, & Castillo-Romero A. (2023). Repurposing Terfenadine as a Novel Antigiardial Compound. Pharmaceuticals, 16(9), 1332.

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Immunostaining of Differentiated PC12 Cells

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After eight days of cells cultured in NGF(+) or NGF(−) Differentiation Medium on different scaffolds, the immunostaining of cytoskeletal proteins

α

-Tubulin was carried out to observe the cell phenotype and neurite extension of differentiated PC12 cells. The reason for choosing α-Tubulin as the staining marker was based on former studies [28 (link),29 (link)]. All the scaffolds were rinsed with PBS, fixed in neutral buffer formalin solution (Sigma) for 20 min and permeabilized with 0.25% Triton X-100. The nonspecific binding was blocked by incubating with 3% BSA in PBS for 90 min. Subsequently the samples were incubated with a neuronal-specific Mouse anti-

α

-Tubulin antibody (Invitrogen, Camarillo, CA, USA), followed by the staining of Alexa Fluor 594 Goat Anti-Mouse IgG (H + L) antibody (Invitrogen), and the nuclei was stained with DAPI (Thermo Scientific). The immunostained samples were mounted on glass slides and fluorescent images were taken using a laser scanning confocal microscope (Zeiss LSM700). Some samples incubated with PBS instead of anti-α-Tubulin antibody were used as the blank control group.

Hu J., Tian L., Prabhakaran M.P., Ding X, & Ramakrishna S. (2016). Fabrication of Nerve Growth Factor Encapsulated Aligned Poly(ε-Caprolactone) Nanofibers and Their Assessment as a Potential Neural Tissue Engineering Scaffold. Polymers, 8(2), 54.

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Cell Treatment and Imaging Protocol

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Polylysine-coated slides were sterilized with 70% ethanol prior to plating with either PC-3 or C4-2B cells. Two hundred microliters of cells at a concentration of 5×10⁵ cells/ml were applied to the slides in an area outlined by a wax pen. Cells were allowed to adhere to the slides overnight while being kept in a 150-mm dish. The following day, medium was aspirated and cells were re-fed with media containing either DMSO vehicle, MMAE (4 nM), or MMAEp (96 nM) for 24 h. Cells were fixed in methanol and extensively washed with phosphate-buffered saline (PBS). The slides were then blocked in goat IgG for 1 h. The slides were probed with mouse anti-α-tubulin antibody (working titer: 0.5 μg/ml, catalogue number 322588, Invitrogen) for 3 h at room temperature and counterstained for 10 minutes with Hoechst stain (0.2 μg/ml) at room temperature. Both stains were applied in the dark with extensive PBS washes in between applications of stain. Cells were imaged using a Nikon Eclipse 80i microscope connected to a Photometrics CoolSnap EZ camera. The image acquisition software utilized for this study was NIS Elements (Nikon Instruments Inc., Melville, NY, USA). Quantification of immunofluorescence signal was accomplished using the CellProfiler program [30 (link),31 (link)].

Cunningham D., Parajuli K.R., Zhang C., Wang G., Mei J., Zhang Q., Liu S, & You Z. (2016). Monomethyl Auristatin E Phosphate Inhibits Human Prostate Cancer Growth. The Prostate, 76(15), 1420-1430.

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