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    Alomone Labs psalmotoxin 1
    D-Tubocurarine (d-TC) induces presynaptic homeostatic potentiation (PHP) and this is blocked by ASIC inhibitors. A. mEPPs and EPPs were measured before and after a 10 min incubation in d-TC (n = 75 NMJs, 15 mice). Quantal Content (QC) was estimated from corrected EPP and mEPP amplitudes. The data are presented as violin plots, which show the probability density of the data at different values. The mean differences are shown as estimation plots (see Experimental Procedure: Statistical Analysis.) The insets are sample traces of EPPs and mEPPs before (black line) and after (grey line) d-TC. Each trace is the average of 10 measurements. Calibration bars indicate 0.2 mv, 2 ms for the mEPPs and 5 mv, 2 ms for the EPPs. B. QC was calculated from corrected mEPP and EPP amplitudes in the presence of 50 μM benzamil (n=25, 5 mice), 100 μM benzamil (n=15, 3 mice), 50 nM <t>PcTx1</t> (n=73, 10 mice), 100 nM PcTx1 (n=25, 5 mice), and 50 nM Mamb3 (n=20, 4 mice) before and after a 10 min incubation in d-TC. For each condition, the ratios of QC after vs. before dTC were calculated and plotted as violin plots. The mean differences are shown as estimation plots. In both A and B, means are depicted as black dots; 95% confidence intervals are indicated by the ends of vertical error bars. In all figures, p values are indicated and were calculated from an ordinary one-way ANOVA.
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    D-Tubocurarine (d-TC) induces presynaptic homeostatic potentiation (PHP) and this is blocked by ASIC inhibitors. A. mEPPs and EPPs were measured before and after a 10 min incubation in d-TC (n = 75 NMJs, 15 mice). Quantal Content (QC) was estimated from corrected EPP and mEPP amplitudes. The data are presented as violin plots, which show the probability density of the data at different values. The mean differences are shown as estimation plots (see Experimental Procedure: Statistical Analysis.) The insets are sample traces of EPPs and mEPPs before (black line) and after (grey line) d-TC. Each trace is the average of 10 measurements. Calibration bars indicate 0.2 mv, 2 ms for the mEPPs and 5 mv, 2 ms for the EPPs. B. QC was calculated from corrected mEPP and EPP amplitudes in the presence of 50 μM benzamil (n=25, 5 mice), 100 μM benzamil (n=15, 3 mice), 50 nM PcTx1 (n=73, 10 mice), 100 nM PcTx1 (n=25, 5 mice), and 50 nM Mamb3 (n=20, 4 mice) before and after a 10 min incubation in d-TC. For each condition, the ratios of QC after vs. before dTC were calculated and plotted as violin plots. The mean differences are shown as estimation plots. In both A and B, means are depicted as black dots; 95% confidence intervals are indicated by the ends of vertical error bars. In all figures, p values are indicated and were calculated from an ordinary one-way ANOVA.

    Journal: Neuroscience

    Article Title: Extracellular Protons Mediate Presynaptic Homeostatic Potentiation at the Mouse Neuromuscular Junction

    doi: 10.1016/j.neuroscience.2021.01.036

    Figure Lengend Snippet: D-Tubocurarine (d-TC) induces presynaptic homeostatic potentiation (PHP) and this is blocked by ASIC inhibitors. A. mEPPs and EPPs were measured before and after a 10 min incubation in d-TC (n = 75 NMJs, 15 mice). Quantal Content (QC) was estimated from corrected EPP and mEPP amplitudes. The data are presented as violin plots, which show the probability density of the data at different values. The mean differences are shown as estimation plots (see Experimental Procedure: Statistical Analysis.) The insets are sample traces of EPPs and mEPPs before (black line) and after (grey line) d-TC. Each trace is the average of 10 measurements. Calibration bars indicate 0.2 mv, 2 ms for the mEPPs and 5 mv, 2 ms for the EPPs. B. QC was calculated from corrected mEPP and EPP amplitudes in the presence of 50 μM benzamil (n=25, 5 mice), 100 μM benzamil (n=15, 3 mice), 50 nM PcTx1 (n=73, 10 mice), 100 nM PcTx1 (n=25, 5 mice), and 50 nM Mamb3 (n=20, 4 mice) before and after a 10 min incubation in d-TC. For each condition, the ratios of QC after vs. before dTC were calculated and plotted as violin plots. The mean differences are shown as estimation plots. In both A and B, means are depicted as black dots; 95% confidence intervals are indicated by the ends of vertical error bars. In all figures, p values are indicated and were calculated from an ordinary one-way ANOVA.

    Article Snippet: In and , benzamil hydrochloride hydrate (Sigma-Aldrich, St. Louis, MO), psalmotoxin-1 (PcTx1; Alomone Labs), or mambalgin-3 (Mamb3; Alomone Labs), were applied (in the presence of μ-Ctx) for 30 minutes prior to recording.

    Techniques: Incubation

    High concentrations of the ASIC inhibitors Benzamil and PcTx1 reduce mEPP and EPP amplitude. MEPPs, EPPs and Quantal Content are plotted from NMJs incubated in either control saline (n=75, same data show in 1A), 100 μM benzamil (n=15, 3 mice), and 100 nM PcTx1 (n=25, 5 mice). Data are plotted as in Fig. 1. P values are indicated and were calculated from Student’s t-test or ordinary one-way ANOVA.

    Journal: Neuroscience

    Article Title: Extracellular Protons Mediate Presynaptic Homeostatic Potentiation at the Mouse Neuromuscular Junction

    doi: 10.1016/j.neuroscience.2021.01.036

    Figure Lengend Snippet: High concentrations of the ASIC inhibitors Benzamil and PcTx1 reduce mEPP and EPP amplitude. MEPPs, EPPs and Quantal Content are plotted from NMJs incubated in either control saline (n=75, same data show in 1A), 100 μM benzamil (n=15, 3 mice), and 100 nM PcTx1 (n=25, 5 mice). Data are plotted as in Fig. 1. P values are indicated and were calculated from Student’s t-test or ordinary one-way ANOVA.

    Article Snippet: In and , benzamil hydrochloride hydrate (Sigma-Aldrich, St. Louis, MO), psalmotoxin-1 (PcTx1; Alomone Labs), or mambalgin-3 (Mamb3; Alomone Labs), were applied (in the presence of μ-Ctx) for 30 minutes prior to recording.

    Techniques: Incubation

    Lowering extracellular pH underlies the upregulation of QC triggered by d-TC. A. mEPPs and EPPs were measured and QC estimated before and after a 10 min incubation in pH 7.2 saline with 100nM PcTx1 present throughout (n=25, 5 mice). The data for mEPPs, EPPs and QC are presented as violin plots. The plot on the far right includes the data from Fig. 3B to depicts the effect of pH 7.2 saline under control conditions vs. in the presence of 100 nM PcTx1. The Mean differences are plotted as a bootstrap sampling distribution. Means are depicted as black dots; 95% confidence intervals are indicated by the ends of vertical error bars. In all figures, p values are indicated and were calculated from an ordinary one-way ANOVA. B. mEPPs and EPPs were measured and QC was estimated in pH 7.4 saline, after a 7 min incubation in pH 7.2 saline, and then after a 15 min incubation with d-TC, still in pH 7.2 saline (n = 5 mice). Although dTC had its normal effects on mEPP and EPP amplitude (compare to Fig. 1A), dTC did not alter QC when compared to pH 7.2 saline. The data is plotted as in panel A. C. The ratio of QC after application of dTC compared to before dTC was applied (post/pre dTC) is shown for pH 7.4 saline (this is the same data presented in Fig 1B, Control) and for pH 7.2 saline (the same data presented in Fig. 4B).

    Journal: Neuroscience

    Article Title: Extracellular Protons Mediate Presynaptic Homeostatic Potentiation at the Mouse Neuromuscular Junction

    doi: 10.1016/j.neuroscience.2021.01.036

    Figure Lengend Snippet: Lowering extracellular pH underlies the upregulation of QC triggered by d-TC. A. mEPPs and EPPs were measured and QC estimated before and after a 10 min incubation in pH 7.2 saline with 100nM PcTx1 present throughout (n=25, 5 mice). The data for mEPPs, EPPs and QC are presented as violin plots. The plot on the far right includes the data from Fig. 3B to depicts the effect of pH 7.2 saline under control conditions vs. in the presence of 100 nM PcTx1. The Mean differences are plotted as a bootstrap sampling distribution. Means are depicted as black dots; 95% confidence intervals are indicated by the ends of vertical error bars. In all figures, p values are indicated and were calculated from an ordinary one-way ANOVA. B. mEPPs and EPPs were measured and QC was estimated in pH 7.4 saline, after a 7 min incubation in pH 7.2 saline, and then after a 15 min incubation with d-TC, still in pH 7.2 saline (n = 5 mice). Although dTC had its normal effects on mEPP and EPP amplitude (compare to Fig. 1A), dTC did not alter QC when compared to pH 7.2 saline. The data is plotted as in panel A. C. The ratio of QC after application of dTC compared to before dTC was applied (post/pre dTC) is shown for pH 7.4 saline (this is the same data presented in Fig 1B, Control) and for pH 7.2 saline (the same data presented in Fig. 4B).

    Article Snippet: In and , benzamil hydrochloride hydrate (Sigma-Aldrich, St. Louis, MO), psalmotoxin-1 (PcTx1; Alomone Labs), or mambalgin-3 (Mamb3; Alomone Labs), were applied (in the presence of μ-Ctx) for 30 minutes prior to recording.

    Techniques: Incubation, Sampling