Monolayers of control or MAG-expressing Chinese hamster ovary (CHO) cells or primary astrocytes were plated on eight well chamber slides as previously described.13 (link) Purified P1 hippocampal, P1 cortical, P5–P6 CGN or P5–P6 DRG rat neurons were diluted to 35 000 cells/ml in Sato’s media and treated with either 1 mM dbcAMP (Calbiochem), mixed population or recombinant Histones (1–20 µg/ml) or with APC (1–20 µg/ml). Neurons were incubated for 14–18 h at 37°C and immunostained using a monoclonal anti-βIII tubulin antibody (Tuj1; Covance) and Alexa Fluor 568-conjugated anti-mouse IgG (Invitrogen). For quantification, images were taken, and the length of the longest neurite for each neuron was measured using MetaMorph software (Molecular Devices).
Anti βiii tubulin antibody tuj1
Manufactured by Fortrea
Anti-βIII tubulin antibody (Tuj1) is a protein-binding antibody that specifically recognizes the βIII isoform of tubulin, a key structural component of microtubules in neurons and other cell types. This antibody is commonly used in research applications as a marker for neuronal differentiation and development.
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2 protocols using anti βiii tubulin antibody tuj1
1
Neurite Outgrowth Assay in Primary Neurons
2
Immunostaining of Neuronal Cultures
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After culturing for 6 days, the DRG neurons
were immunostained with anti-βIII tubulin antibody (Tuj1) (Covance,
San Diego, CA). Briefly, the samples were fixed and blocked as described
above and then incubated with antibody diluted (1:500) in PBS that
contained 2% NGS overnight at 4 °C. The Tuj1 marker was detected
using AlexaFluor 488 goat anti-mouse IgG (1:200; Invitrogen) secondary
antibody and imaged as described above.
were immunostained with anti-βIII tubulin antibody (Tuj1) (Covance,
San Diego, CA). Briefly, the samples were fixed and blocked as described
above and then incubated with antibody diluted (1:500) in PBS that
contained 2% NGS overnight at 4 °C. The Tuj1 marker was detected
using AlexaFluor 488 goat anti-mouse IgG (1:200; Invitrogen) secondary
antibody and imaged as described above.
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