rabbit polyclonal anti na v 1 6  (Alomone Labs)

Journal: eLife

Article Title: Calpain fosters the hyperexcitability of motoneurons after spinal cord injury and leads to spasticity

doi: 10.7554/eLife.51404

Figure Lengend Snippet:

Article Snippet: Antibody , rabbit polyclonal anti-Na v 1.6 , Alomone , Cat # ASC-009 RRID: AB_2040202 , (1:200).

Techniques: Diagnostic Assay, Software, Microscopy, Imaging

Journal: Scientific Reports

Article Title: Amyloid β-Induced Upregulation of Na v 1.6 Underlies Neuronal Hyperactivity in Tg2576 Alzheimer’s Disease Mouse Model

doi: 10.1038/s41598-019-50018-1

Figure Lengend Snippet: Effect of Aβ 1–42 exposure on Na V 1.6 protein expression and activity in primary hippocampal neurons at 10–12 DIV. ( A ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.1 protein expression in primary hippocampal neurons under control conditions and after 5 μM Aβ 1–42 (24 h). Values are expressed as mean ± SEM of 3 independent experimental sessions. ( B ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.2 protein expression in primary hippocampal neurons under control conditions and after 5 μM Aβ 1–42 (24 h). Values are expressed as mean ± SEM of 3 independent experimental sessions. ( C ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.6 protein expression in primary hippocampal neurons under control conditions and after 5 μM Aβ 1–42 (24 h). Values are expressed as mean ± SEM of 3 independent experimental sessions. ** p < 0.01 versus control. ( D ) Representative traces of Na + currents recorded under control conditions, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min) in primary hippocampal neurons. (E) Representative traces of Na + currents recorded after 5 μM Aβ 1–42 (24 h) alone, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min) in primary hippocampal neurons. ( F ) Normalization of Na + current densities, at −20 mV, represented in panel D and E. The number of cells used for each experimental condition is noted on the bars, values are expressed as percentage mean ± SEM of 3 independent experimental sessions. *** p < 0.001 versus control, # p < 0.001 versus control Aβ 1–42 . (G) Representative western blot of Na V 1.6 protein expression (top) in the presence of siNa V 1.6 (50 nM; 48 h) in primary hippocampal neurons at 12 DIV. Quantification of siNav1.6 inhibition in primary hippocampal neurons (bottom). Values are expressed as percentage mean ± SEM of 3 independent experimental sessions. ** p < 0.01 versus control neurons.

Article Snippet: In brief, cell cultures were fixed in 4% paraformaldehyde in PBS for 30 min. After blockage with Rodent M Block (Biocare Medical, Concord, CA, USA) for 1 hour, cells were incubated with rabbit polyclonal anti-Na V 1.6 antibody (1:2000 Alomone Labs, Israel) and mouse monoclonal anti-MAP2 antibody (1:2000, Sigma Aldrich, Milan, Italy) at 4 °C overnight for 24 hours.

Techniques: Expressing, Activity Assay, Western Blot, Inhibition

Journal: Scientific Reports

Article Title: Amyloid β-Induced Upregulation of Na v 1.6 Underlies Neuronal Hyperactivity in Tg2576 Alzheimer’s Disease Mouse Model

doi: 10.1038/s41598-019-50018-1

Figure Lengend Snippet: Expression and activity of Na V 1.6 channels in Tg2576 primary hippocampal neurons. ( A ) Representative traces of Na + currents recorded in WT and Tg2576 primary hippocampal neurons after 8 and 12 DIV. ( B ) Normalization of Na + current densities at −20 mV represented in panel A. Values are expressed as mean ± SEM of current densities of 3 independent experimental sessions. The number of cells used for each experimental condition is noted on the bars, values are expressed as percentage mean ± SEM of 3 independent experimental sessions. *** p < 0.001 versus WT. ( C ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.1 protein expression in WT and Tg2576 primary hippocampal neurons after 12 DIV. Values are expressed as mean ± SEM of 3 independent experimental sessions. ( D ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.2 protein expression in WT and Tg2576 primary hippocampal neurons after 12 DIV. Values are expressed as mean ± SEM of 3 independent experimental sessions. ( E ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.6 protein expression in WT and Tg2576 primary hippocampal neurons after 12 DIV. Values are expressed as mean ± SEM of 3 independent experimental sessions. ** p < 0.01 versus WT. (F) Representative confocal images displaying Na V 1.6 distribution in WT (left) and Tg2576 (right) primary hippocampal neurons after 12 DIV. Scale bars: 20 μm.

Article Snippet: In brief, cell cultures were fixed in 4% paraformaldehyde in PBS for 30 min. After blockage with Rodent M Block (Biocare Medical, Concord, CA, USA) for 1 hour, cells were incubated with rabbit polyclonal anti-Na V 1.6 antibody (1:2000 Alomone Labs, Israel) and mouse monoclonal anti-MAP2 antibody (1:2000, Sigma Aldrich, Milan, Italy) at 4 °C overnight for 24 hours.

Techniques: Expressing, Activity Assay, Western Blot

Journal: Scientific Reports

Article Title: Amyloid β-Induced Upregulation of Na v 1.6 Underlies Neuronal Hyperactivity in Tg2576 Alzheimer’s Disease Mouse Model

doi: 10.1038/s41598-019-50018-1

Figure Lengend Snippet: Effect of siNa V 1.6 and anisomycin on Na V 1.6 protein expression and activity in Tg2576 primary hippocampal neurons. ( A ) Representative traces of Na + currents recorded in WT primary hippocampal neurons after 12 DIV under control conditions, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min). ( B ) Representative traces of Na + currents recorded in Tg2576 primary hippocampal neurons after 12 DIV under control conditions, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min). ( C ) Normalization of Na + current densities at −20 mV represented in panel A and B. The number of cells used for each experimental condition is noted on the bars, values are expressed as percentage mean ± SEM of 3 independent experimental sessions. ** p < 0.01 versus control WT, *** p < 0.001 versus control WT, # p < 0.001 versus control Tg2576. ( D ) Representative current tracings recorded in the gap-free mode in WT and Tg2576 hippocampal neurons after 12 DIV under control conditions, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min). ( E ) Quantification of spike frequency recorded in WT and Tg2576 hippocampal neurons after 12 DIV under control conditions, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min). The number of cells used for each experimental condition is noted on the bars, values are expressed as percentage mean ± SEM of 3 independent experimental sessions. *** p < 0.001 versus WT. # p < 0.001 versus control Tg2576. ( F ) Quantification of membrane depolarization recorded in WT and Tg2576 primary hippocampal neurons after 12 DIV under control conditions, in the presence of siNa V 1.6 (50 nM; 48 h) and in the presence of anisomycin (10 μM; 30 min). The number of cells used for each experimental condition is noted on the bars, values are expressed as percentage mean ± SEM of 3 independent experimental sessions. ** p < 0.01 versus WT. # p < 0.001 versus control Tg2576

Article Snippet: In brief, cell cultures were fixed in 4% paraformaldehyde in PBS for 30 min. After blockage with Rodent M Block (Biocare Medical, Concord, CA, USA) for 1 hour, cells were incubated with rabbit polyclonal anti-Na V 1.6 antibody (1:2000 Alomone Labs, Israel) and mouse monoclonal anti-MAP2 antibody (1:2000, Sigma Aldrich, Milan, Italy) at 4 °C overnight for 24 hours.

Techniques: Expressing, Activity Assay

Journal: Scientific Reports

Article Title: Amyloid β-Induced Upregulation of Na v 1.6 Underlies Neuronal Hyperactivity in Tg2576 Alzheimer’s Disease Mouse Model

doi: 10.1038/s41598-019-50018-1

Figure Lengend Snippet: Immunocytochemical analysis of Na V 1.6 protein expression after anisomycin treatment in Tg2576 primary hippocampal neurons at 12 DIV. ( A ) Confocal double immunofluorescence images displaying Na V 1.6 (green) and MAP2 (red) distribution in WT (a-c) and Tg2576 primary hippocampal neurons in the absence (d-f) or in the presence (g-i) of anisomycin. Scale bars in a-i: 20 μm. ( B ) Quantitative analyses of Na V 1.6-positive puncta within the soma of WT and Tg2576 primary hippocampal neurons in the absence or in the presence of anisomycin. Scale bars: 5 μm. Data are expressed as mean ± SEM of values obtained from 20 cells per group in 3 independent experimental sessions. ** p < 0.01 versus WT; # p < 0.001 versus Tg2576.

Article Snippet: In brief, cell cultures were fixed in 4% paraformaldehyde in PBS for 30 min. After blockage with Rodent M Block (Biocare Medical, Concord, CA, USA) for 1 hour, cells were incubated with rabbit polyclonal anti-Na V 1.6 antibody (1:2000 Alomone Labs, Israel) and mouse monoclonal anti-MAP2 antibody (1:2000, Sigma Aldrich, Milan, Italy) at 4 °C overnight for 24 hours.

Techniques: Expressing, Immunofluorescence

Journal: Scientific Reports

Article Title: Amyloid β-Induced Upregulation of Na v 1.6 Underlies Neuronal Hyperactivity in Tg2576 Alzheimer’s Disease Mouse Model

doi: 10.1038/s41598-019-50018-1

Figure Lengend Snippet: Evaluation of Na V 1.6 protein expression in the hippocampus of 3-month-old WT and Tg2576 mice. ( A ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.1 protein expression in the hippocampus of WT and Tg2576 mice. Values are expressed as mean ± SEM of 3 independent experimental sessions. ( B ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.2 protein expression in the hippocampus of WT and Tg2576 mice. Values are expressed as mean ± SEM of 3 independent experimental sessions. ( C ) Representative western blot (top) and densitometric quantification (bottom) of Na V 1.6 protein expression in the hippocampus of WT and Tg2576 mice. Values are expressed as mean ± SEM of 3 independent experimental sessions. ** p < 0.01 versus WT. ( D ) Confocal double immunofluorescence images displaying Na V 1.6 (green) and MAP2 (red) distribution in the hippocampus of 3-month-old WT (a-c) and Tg2576 mice (d-f). Scale bars in a-f: 20 μm. (E) Quantitative analyses of Na V 1.6-positive puncta within the soma of WT and Tg2576 neurons in the hippocampus of 3-month-old mice. Values are expressed as mean ± SEM of 3 independent experimental sessions. **p < 0.01 versus WT.

Article Snippet: In brief, cell cultures were fixed in 4% paraformaldehyde in PBS for 30 min. After blockage with Rodent M Block (Biocare Medical, Concord, CA, USA) for 1 hour, cells were incubated with rabbit polyclonal anti-Na V 1.6 antibody (1:2000 Alomone Labs, Israel) and mouse monoclonal anti-MAP2 antibody (1:2000, Sigma Aldrich, Milan, Italy) at 4 °C overnight for 24 hours.

Techniques: Expressing, Western Blot, Immunofluorescence