polyclonal asic2 antibody  (Alomone Labs)

Journal: Cell Death and Differentiation

Article Title: Acidotoxicity and acid-sensing ion channels contribute to motoneuron degeneration

doi: 10.1038/cdd.2012.158

Figure Lengend Snippet: Expression of asic1, asic2 and asic3 during ALS disease progression. (a) Co-staining of lumbar spinal cords from tg SOD1- and asic1a-deficient mice with anti-ASIC1/ASIC2 (red) and anti-NeuN/GFAP (green) antibodies. Arrowheads show co-localization of ASIC1 and ASIC2 with NeuN on cell soma of neurons with motoneuron morphology. Scale bar=20 μm. (b) asic1, −2 and −3 mRNA expression in the transgenic (tg) SOD1 mouse; lumbar spinal cord lysates show a significant upregulation of the asic1a and asic2a mRNA; *P<0.01. Data displayed as mean±S.D.; n=6/group. (c) Co-staining of lumbar spinal cords across disease progression in the tg SOD1 mouse with anti-ASIC1/ASIC2 (red) and anti-NeuN/smi-32 (green) antibodies. Co-localization of ASIC antibodies with motoneurons is indicated by arrowheads. Scale bar=15 μm. (d) An increase in ASIC1 and ASIC2 protein levels in the tg SOD1 mouse; *P<0.01. Data displayed as mean±S.D.; n=6/group. (e) A representative image of ASIC1 and ASIC2 protein (double band) levels by western blotting, n=3 experiments with tubulin as loading control. (f) ASIC2 immunohistochemistry in ventral motoneurons of lumbar spinal cord sections from ALS and non-ALS patients. n=3/group. Scale bar=50 μm

Article Snippet: Sections were incubated with a polyclonal ASIC2 antibody (1 : 200, Alomone Labs, Jerusalem, Israel), biotinylated secondary antibody (1 : 500; Jackson Laboratories) and visualized using Sigma fast 3,3-diaminobenzadine tablets.

Techniques: Expressing, Staining, Transgenic Assay, Western Blot, Immunohistochemistry

Journal: Cell Death and Differentiation

Article Title: Acidotoxicity and acid-sensing ion channels contribute to motoneuron degeneration

doi: 10.1038/cdd.2012.158

Figure Lengend Snippet: (A) Characteristics of cases and controls used from ALS and non-ALS patients. (B) Mean staining intensity values for ASIC2 protein in ALS and non-ALS patients

Article Snippet: Sections were incubated with a polyclonal ASIC2 antibody (1 : 200, Alomone Labs, Jerusalem, Israel), biotinylated secondary antibody (1 : 500; Jackson Laboratories) and visualized using Sigma fast 3,3-diaminobenzadine tablets.

Techniques: Staining

Journal: Scientific Reports

Article Title: Syntabulin regulates the trafficking of PICK1-containing vesicles in neurons

doi: 10.1038/srep20924

Figure Lengend Snippet: ( A ) Neurons were infected with syntabulin shRNA lentivirus as indicated at DIV5 and examined at DIV13. Syntabulin knockdown significantly increased total ASIC2 expression but reduced surface ASIC2 levels as compared with the levels in control and scrambled-shRNA control. ( B ) Quantification of normalized ASIC2/GAPDH ratio, total ASIC2/surface ASIC2 ratio and surface ASIC2 levels. One-way ANOVA and Sidak test; ** P = 0.008, and P = 0.009 for Stb sh#1 Total ASIC2/GAPDH and Surface ASIC2/GAPDH compared to control respectively; * P = 0.044 and * P = 0.016 for Stb #1 total ASIC2/GAPDH and Surface/Total ASIC2 ratio compared to scrambled shRNA control respectively; * P = 0.020 for Stb #1 Surface/Total ASIC2 ratio compared to control; “ns” P > 0.05. n = 4 − 6 independent experiments. Error bars represent the SEM. ( C , D ) Hoechst staining of neurons after syntabulin knockdown, in pH 7.4 solution and pH6.0 solution. Knockdown of syntabulin significantly increased cell death as compared with controls in pH6.0. Scale bar = 20 μm. ( E ) Quantification of normalized total Hoechst intensity. One-way ANOVA and Sidak test; * P = 0.035 compared to control; * P = 0.043 compared to scrambled-shRNA control; “ns” P > 0.05; n = 6 independent experimental repeats. Error bars represent the SEM.

Article Snippet: The following antibodies were purchased: rabbit polyclonal ASIC2 antibody, Proteintech (17851-1-AP); mouse myc and β-tubulin antibodies, DSHB (9E10 and E7); mouse GAPDH antibody, Beyotime (GA019); mouse Map2 antibody, Sigma (M9942); and mouse β-actin antibody, Sigma (A5316).

Techniques: Infection, shRNA, Knockdown, Expressing, Control, Staining

Journal: Cell Death and Differentiation

Article Title: Acidotoxicity and acid-sensing ion channels contribute to motoneuron degeneration

doi: 10.1038/cdd.2012.158

Figure Lengend Snippet: Expression of asic1, asic2 and asic3 during ALS disease progression. (a) Co-staining of lumbar spinal cords from tg SOD1- and asic1a-deficient mice with anti-ASIC1/ASIC2 (red) and anti-NeuN/GFAP (green) antibodies. Arrowheads show co-localization of ASIC1 and ASIC2 with NeuN on cell soma of neurons with motoneuron morphology. Scale bar=20 μm. (b) asic1, −2 and −3 mRNA expression in the transgenic (tg) SOD1 mouse; lumbar spinal cord lysates show a significant upregulation of the asic1a and asic2a mRNA; *P<0.01. Data displayed as mean±S.D.; n=6/group. (c) Co-staining of lumbar spinal cords across disease progression in the tg SOD1 mouse with anti-ASIC1/ASIC2 (red) and anti-NeuN/smi-32 (green) antibodies. Co-localization of ASIC antibodies with motoneurons is indicated by arrowheads. Scale bar=15 μm. (d) An increase in ASIC1 and ASIC2 protein levels in the tg SOD1 mouse; *P<0.01. Data displayed as mean±S.D.; n=6/group. (e) A representative image of ASIC1 and ASIC2 protein (double band) levels by western blotting, n=3 experiments with tubulin as loading control. (f) ASIC2 immunohistochemistry in ventral motoneurons of lumbar spinal cord sections from ALS and non-ALS patients. n=3/group. Scale bar=50 μm

Article Snippet: Membranes were incubated with the following primary antibodies: polyclonal ASIC1 antibody (1 : 1000, a kind gift from Dr. John Wemmie), polyclonal ASIC2 antibody (1 : 500, Novus Biologicals, Colorado, USA), polyclonal SOD1 antibody (1 : 500 Sigma-Aldrich, Dublin, Ireland), monoclonal tubulin antibody (1 : 5000, Sigma-Aldrich, Dublin, Ireland) and polyclonal actin antibody (1 : 5000, Abcam, Sussex, UK).

Techniques: Expressing, Biomarker Discovery, Staining, Transgenic Assay, Western Blot, Control, Immunohistochemistry

Journal: Cell Death and Differentiation

Article Title: Acidotoxicity and acid-sensing ion channels contribute to motoneuron degeneration

doi: 10.1038/cdd.2012.158

Figure Lengend Snippet: (A) Characteristics of cases and controls used from ALS and non-ALS patients. (B) Mean staining intensity values for ASIC2 protein in ALS and non-ALS patients

Article Snippet: Membranes were incubated with the following primary antibodies: polyclonal ASIC1 antibody (1 : 1000, a kind gift from Dr. John Wemmie), polyclonal ASIC2 antibody (1 : 500, Novus Biologicals, Colorado, USA), polyclonal SOD1 antibody (1 : 500 Sigma-Aldrich, Dublin, Ireland), monoclonal tubulin antibody (1 : 5000, Sigma-Aldrich, Dublin, Ireland) and polyclonal actin antibody (1 : 5000, Abcam, Sussex, UK).

Techniques: Staining, Biomarker Discovery, Control