The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml.
Use a concentration of 2 µg/ml.
Use a concentration of 5 µg/ml.
Use 1µg for 106 cells. ab91545-Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
FunctionTubulin 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.
Tissue specificityUbiquitously expressed with highest levels in spleen, thymus and immature brain.
Involvement in diseaseCortical dysplasia, complex, with other brain malformations 6 Skin creases, congenital symmetric circumferential, 1
Sequence similaritiesBelongs to the tubulin family.
DomainThe highly acidic C-terminal region may bind cations such as calcium.
Post-translational modificationsSome glutamate residues at the C-terminus are polyglutamylated, resulting in polyglutamate chains on the gamma-carboxyl group (PubMed:26875866). Polyglutamylation plays a key role in microtubule severing by spastin (SPAST). SPAST preferentially recognizes and acts on microtubules decorated with short polyglutamate tails: severing activity by SPAST increases as the number of glutamates per tubulin rises from one to eight, but decreases beyond this glutamylation threshold (PubMed:26875866). Some glutamate residues at the C-terminus are 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). Both polyglutamylation and monoglycylation can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of monoglycylation is still unclear. Phosphorylated on Ser-172 by CDK1 during the cell cycle, from metaphase to telophase, but not in interphase. This phosphorylation inhibits tubulin incorporation into microtubules.
Immunocytochemistry/Immunofluorescence analysis of 3T3 mouse embryonal fibroblast cells labelling beta Tubulin with ab7792 at 2 μg/mL. A Cy®5 conjugated goat anti-mouse IgM was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Overlay histogram showing HeLa cells stained with ab7792 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100® 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 (ab7792, 1µg/1x10^6 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100® used under the same conditions.
References for Anti-beta Tubulin antibody [TU-06] (ab7792)
This product has been referenced in:
Staples CJ et al. MRNIP/C5orf45 Interacts with the MRN Complex and Contributes to the DNA Damage Response. Cell Rep16:2565-75 (2016).
Read more (PubMed: 27568553) »
McNeill E et al. Regulation of iNOS function and cellular redox state by macrophage Gch1 reveals specific requirements for tetrahydrobiopterin in NRF2 activation. Free Radic Biol Med79:206-16 (2015).
Read more (PubMed: 25451639) »