The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 5.1µl for 106 cells.
ab106163 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 1 µg/ml.
Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain.
Belongs to the tubulin family.
Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules. Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
ICC/IF image of ab64503 stained human HeLa cells. The cells were methanol fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab64503, 1µg/ml, FITC conjugated (green)) for 1h at room temperature. 1% BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry (Frozen sections) - Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (FITC) (ab64503)This image is a courtesy of Abreview submitted by Anonymous
ab64503 staining alpha Tubulin in Xenopus laevis stage 36, transverse cryosection tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with MEMFa and blocked with 2% BSA/Serum for 1 hour at 230C. The sample was incubated with primary antibody (1/100) for 1 hour at 230C. No secondary antibody was used. FITC-tubulin shown in green highlights neurons in neural tube and ciliated epidermal cells. DAPI shows staining in grey. Laminin (Abcam`s ab11575) is in blue (donkey anti-rabbit Cy5 secondary). E-cadherin is in red (donkey anti-mouse Cy3 secondary).
Overlay histogram showing HeLa cells stained with ab64503 (red line). The cells were fixed with 80% methanol (5 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 (ab64503, 0.5µg/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 (1µg/1x106 cells ). Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.