antineuronal specific βiii tubulin  (R&D Systems)

Journal: The Journal of Biological Chemistry

Article Title: ABCA13 dysfunction associated with psychiatric disorders causes impaired cholesterol trafficking

doi: 10.1074/jbc.RA120.015997

Figure Lengend Snippet: Full-size ABCA13 is localized in intracellular vesicles. A , Western blot analysis of kidneys from WT mice using anti-ABCA13 antibody. β-actin was used as a loading control. B , HEK293 cells were transiently transfected with mouse ABCA13. ABCA13 expression was confirmed by western blotting. β-actin was used as a loading control. C , cells were immunostained with anti-ABCA13 antibody ( green ) and anti-β-tubulin antibody ( magenta ) after fixation and permeabilization. Nuclei were stained with TOTO-3 ( blue ). Scale bars represent 10 μm.

Article Snippet: Mouse antineuron specific βIII-tubulin antibody was purchased from Merck Millipore.

Techniques: Western Blot, Control, Transfection, Expressing, Staining

Journal: The Journal of Biological Chemistry

Article Title: ABCA13 dysfunction associated with psychiatric disorders causes impaired cholesterol trafficking

doi: 10.1074/jbc.RA120.015997

Figure Lengend Snippet: ABCA13 deletion impairs vesicular cholesterol accumulation and synaptic vesicle endocytosis in neurons. A , primary mouse cortical neurons were treated with 5 mM MβCD for 5 min at 37 °C. Then, the cells were labeled with anti-βIII-tubulin antibody ( magenta ) and the fluorescent probe EGFP-D4 ( green ) after fixation and permeabilization. B , the relative fluorescence intensities of EGFP-D4 signals in individual cells were quantified using ImageJ and shown as means + S.E.M. (n = 30). C , primary mouse cortical neurons were stained with FM 4-64 for 3 min in high K+ buffer. Then, the cells were washed for 10 min in Ca 2+ -free buffer and observed using fluorescence microscopy. D , the relative fluorescence intensities of FM 4-64 signals in individual cells were quantified using ImageJ and shown as means + S.E.M. (n = 30). Scale bars represent 20 μm. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Mouse antineuron specific βIII-tubulin antibody was purchased from Merck Millipore.

Techniques: Labeling, Fluorescence, Staining, Microscopy

Journal: Cerebral Cortex (New York, NY)

Article Title: Neuropilin 1-Sema Signaling Regulates Crossing of Cingulate Pioneering Axons during Development of the Corpus Callosum

doi: 10.1093/cercor/bhp027

Figure Lengend Snippet: Sema3C attracts cingulate cortex neurites in vitro. Cingulate cortex explants grown next to mock-transfected cell blocks (A) or Sema3C-expressing cell blocks (B) for 48 h. Explants have been stained with antineuronal-specific βIII tubulin to visualize neurites. There is no significant difference in neurite outgrowth from explants from either group (C). However, the GR (defined as [(proximal neurite pixels − distal neurite pixels)/(total neurite pixels)) was significantly greater in explants grown next to Sema3C-expressing cell blocks in comparison to controls (*P < 0.0005), indicating that Sema3C acts as an attractant for neurites from cingulate cortex explants. The arrows in A and B indicate the direction of the cell block relative to the explant. Scale bar: 200 μm.

Article Snippet: For the Sema3C culture assay, the primary antibody used was antineuronal-specific βIII tubulin (TuJ-1 clone, R&D Systems, Minneapolis, MN; 1/1000).

Techniques: In Vitro, Transfection, Expressing, Staining, Comparison, Blocking Assay