The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Predicted molecular weight: 50 kDa.
Use a concentration of 5 µg/ml.
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. TUBB3 plays a critical role in proper axon guidance and mantainance.
Tissue specificityExpression is primarily restricted to central and peripheral nervous system.
Involvement in diseaseDefects in TUBB3 are the cause of congenital fibrosis of extraocular muscles type 3A (CFEOM3A) [MIM:600638]. A congenital ocular motility disorder marked by restrictive ophthalmoplegia affecting extraocular muscles innervated by the oculomotor and/or trochlear nerves. It is clinically characterized by anchoring of the eyes in downward gaze, ptosis, and backward tilt of the head. Congenital fibrosis of extraocular muscles type 3 presents as a non-progressive, autosomal dominant disorder with variable expression. Patients may be bilaterally or unilaterally affected, and their oculo-motility defects range from complete ophthalmoplegia (with the eyes fixed in a hypo- and exotropic position), to mild asymptomatic restrictions of ocular movement. Ptosis, refractive error, amblyopia, and compensatory head positions are associated with the more severe forms of the disorder. In some cases the ocular phenotype is accompanied by additional features including developmental delay, corpus callosum agenesis, basal ganglia dysmorphism, facial weakness, polyneuropathy.
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. 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.
Immunoprecipitation - Anti-beta III Tubulin antibody (ab76287)
beta III Tubulin was immunoprecipitated using 0.5mg Mouse Brain tissue, 5µg of Rabbit polyclonal to beta III Tubulin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain tissue lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab76287.
non specific - as present in control (lane 2); We are confident this was due to slight lane contamination and the band seen in the IP lane is our target of interest.
Immunocytochemistry/ Immunofluorescence - beta III Tubulin antibody - C-terminal (ab76287)
ab76287, at a 1/200 dilution, staining bea III Tubulin in chick dorsal root ganglion neurons by Immunoflurescence.
Western blot - beta III Tubulin antibody - C-terminal (ab76287)
All lanes : Anti-beta III Tubulin antibody (ab76287) at 1/2000 dilution
Lane 1 : mouse brain lysates Lane 2 : mouse brain lysates with unphosphorylated beta III Tublin (Ser-444) peptide Lane 3 : mouse brain lysates with phospho beta III Tublin (Ser-444) peptide
Predicted band size : 50 kDa Observed band size : 50 kDa
References for Anti-beta III Tubulin antibody (ab76287)
This product has been referenced in:
Joseph JV et al. Serum-Induced Differentiation of Glioblastoma Neurospheres Leads to Enhanced Migration/Invasion Capacity That Is Associated with Increased MMP9. PLoS One10:e0145393 (2015).
Read more (PubMed: 26700636) »
Joseph JV et al. TGF-ß is an inducer of ZEB1-dependent mesenchymal transdifferentiation in glioblastoma that is associated with tumor invasion. Cell Death Dis5:e1443 (2014).
Read more (PubMed: 25275602) »