mambalgin 3  (Alomone Labs)


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    Alomone Labs mambalgin 3
    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of <t>Mambalgin-3</t> to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Mambalgin 3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms"

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2022.982689

    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Figure Legend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Techniques Used: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.
    Figure Legend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Techniques Used: Concentration Assay, Inhibition

    2) Product Images from "Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms"

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2022.982689

    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Figure Legend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Techniques Used: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.
    Figure Legend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Techniques Used: Concentration Assay, Inhibition

    3) Product Images from "Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms"

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2022.982689

    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Figure Legend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Techniques Used: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.
    Figure Legend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Techniques Used: Concentration Assay, Inhibition

    4) Product Images from "Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms"

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2022.982689

    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Figure Legend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Techniques Used: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.
    Figure Legend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Techniques Used: Concentration Assay, Inhibition

    5) Product Images from "Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms"

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2022.982689

    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Figure Legend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Techniques Used: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.
    Figure Legend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Techniques Used: Concentration Assay, Inhibition

    6) Product Images from "Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms"

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2022.982689

    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Figure Legend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Techniques Used: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.
    Figure Legend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Techniques Used: Concentration Assay, Inhibition

    7) Product Images from "Extracellular Protons Mediate Presynaptic Homeostatic Potentiation at the Mouse Neuromuscular Junction"

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

    Journal: Neuroscience

    doi: 10.1016/j.neuroscience.2021.01.036

    D-Tubocurarine (d-TC) induces presynaptic homeostatic potentiation (PHP) and this is blocked by ASIC inhibitors. A. .) 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.
    Figure Legend Snippet: D-Tubocurarine (d-TC) induces presynaptic homeostatic potentiation (PHP) and this is blocked by ASIC inhibitors. A. .) 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.

    Techniques Used: Mouse Assay, Incubation

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    Alomone Labs mambalgin 3
    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of <t>Mambalgin-3</t> to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.
    Mambalgin 3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    doi: 10.3389/fnmol.2022.982689

    Figure Lengend Snippet: Pharmacology of endogenous proton-activated currents in HEK cells. (A) Individual % inhibition values plotted against concentration of PcTx1 applied during QPatch 48 recordings, with mean shown by horizontal black lines. Data was obtained from cumulative IC 50 assays where each cell was exposed to three concentrations of PcTx1, typically in half log unit increments. (B) Potency of Mambalgin-3 to inhibit pH-activated currents plotted as a cumulative concentration-response IC 50 curve. Symbols are mean ± S.D ( n = 2 or 3). (C) Exemplar current traces from a HEK cell in response to pH 6.5 activation before and after pre-incubation in increasing concentrations of Mambalgin-3. Scale bar is 500 pA and 1 s. (D) Inhibition of proton-activated currents in HEK cells by Amiloride. Data was obtained in a similar fashion to C, with each cell stimulated by pH 6.5 solution and then pre-incubated in increasing concentrations of Amiloride to construct a cumulative mini-IC 50 curve. Symbols are mean ± S.D ( n = 3). (E) Endogenous proton-activated currents are insensitive to Ruthenium Red (RR, 1 μM) but completely inhibited by a non-selective concentration of the ASIC/ENaC/Degenerin antagonist Amiloride (100 μM). The HEK cell was stimulated by pH 6.5 solution and then pre-incubated (at pH 7.4) in RR or Amiloride followed by consecutive stimulation with pH 6.5 solution containing RR or Amiloride.

    Article Snippet: The endogenous proton-activated currents in HEK cells were effectively inhibited by Mambalgin-3, with ∼75% block at 300 nM and a cumulative IC50 value of 119 nM on the QPatch 48 ( ).

    Techniques: Inhibition, Concentration Assay, Activation Assay, Incubation, Construct

    Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Development of ASIC1a ligand-gated ion channel drug screening assays across multiple automated patch clamp platforms

    doi: 10.3389/fnmol.2022.982689

    Figure Lengend Snippet: Validation of a multi-hole ASIC1a assay on QPatch 48. (A) Comparison of efficiency and success rate of single vs. multi-hole QPlate assay format. Mean (± S.D) data for the percentage of wells that passed various quality control (QC) criteria during and after whole-cell recordings in single hole ( n = 4 plates, black squares) or multi-hole mode ( n = 5 plates, filled circles). (B) ASIC1a assay pharmacology in multi-hole mode. Mean (± S.D) concentration-response data for inhibition of ASIC1a currents by Mambalgin-3 ( n = 12), Amiloride ( n = 8) and Benzamil ( n = 4) in multi-hole recordings, with indicated IC 50 and Hill slope values.

    Article Snippet: The endogenous proton-activated currents in HEK cells were effectively inhibited by Mambalgin-3, with ∼75% block at 300 nM and a cumulative IC50 value of 119 nM on the QPatch 48 ( ).

    Techniques: Concentration Assay, Inhibition