Beta tubulin III, also known as Tuj-1, is a class III member of the beta tubulin protein family. Beta tubulins are one of two structural components that form our microtubule network. While general tubulins play a role in a wide range of cellular processes (mitosis, motility, etc) beta tubulin III is specifically localized to neurons. In particular, beta III tubulin’s expression correlates with the earliest phases of neuronal differentiation. For this reason, beta tubulin III has implications in neurogenesis, axon guidance and maintenance. Since it was discovered that the human brain generates new neurons from stem cell pools, beta tubulin III has been used as a marker of positive neuronal identity in many research studies. The following articles use a beta III tubulin antibody in their research of embryonic stem cell and neural progenitors.
beta ‑III Tubulin and Nestin were detected in rat cortical stem cells (Catalog # NSC001) using 5 µg/mL neuron-specific Mouse Anti-Neuron-specific beta ‑III Tubulin Monoclonal Antibody (Catalog # MAB1195) and 10 µg/mL Rat Nestin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2736). Cells were incubated with primary antibodies for 3 hours at room temperature. Cells were stained for beta‑III Tubulin using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and for Nestin using the NorthernLights 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003). Tissue was counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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The first article uses a beta III tubulin antibody in their study of ERK2 and its potential activity in suppressing the self-renewal capacity of embryonic stem cells. The ERK2 protein is dominantly expressed in embryonic stem cells (ES cells), and its inhibition results in hyper phosphorylation of ERK1 and a lack of interaction with the fibroblast growth factor (FGF) pathway, interrupting lineage specification. To begin, Hamilton et al targeted the ERK2 gene in mice, established that ERK2 depletion affects FGF-ERK signaling, and determined that germ layer differentiation in ES is ERK2 independent. Next, they used a beta III tubulin antibody in immunocytochemistry on wild type versus embryo derived Erk2-/- embryonic stem cells to investigate the requirement of ERK2 during neural development. While ERK does suppress self-renewal in ES cells, it was not found to be required for neural cell development or differentiation.
The next article from Haque et al used a beta III tubulin antibody while looking at an engineered N-Cadherin substrate for normal functioning of pluripotent stem cell derived neural progenitor cells. The need for this mechanism is based on the need of better stem cell based treatments for neurodegenerative disease, where production of homogenous populations of neurons is required. For starters, Haque et al established that Akt activation by N-cadherin induces neuronal differentiation. Next, they used a beta III tubulin antibody to illustrate that neuronal differentiation on an N-cadherin substrate can occur without the presence of growth factors. Finally, the Rho/ROCK signaling pathway was suppressed in order to confirm its activity in the N-cadherin signaling pathway as it relates to neurite outgrowth. They found that neurite outgrowth was decreased alongside this suppression after evaluating bright field images and beta III tubulin staining to mark neurite outgrowth.
Novus Biologicals offers beta III tubulin reagents for your research needs including: