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Bio-Rad acrylamide tris hcl ready sds page gels
Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by <t>SDS-PAGE</t> analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM <t>Hepes-Tris,</t> pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.
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1) Product Images from "Interaction of local anesthetics with the K+ channel pore domain"

Article Title: Interaction of local anesthetics with the K+ channel pore domain

Journal: Channels

doi: 10.4161/chan.24455

Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by SDS-PAGE analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM Hepes-Tris, pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.
Figure Legend Snippet: Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by SDS-PAGE analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM Hepes-Tris, pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.

Techniques Used: SDS Page, Incubation

Figure 2. Destabilization of KcsA tetramer as function of LA concentration. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing concentrations of lidocaine (●) or tetracaine (△, □ ). The sample assay mixture contained 10 mM Hepes-Tris, pH 7.5, 100 mM cholineCl and either 5 mM KCl (● , △) or no added KCl (□ ). Samples were incubated either at 90°C (● , △) or 22 °C (□ ) for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of LA concentration as described in Materials and Methods . Fit parameters: lidocaine, 5 mM KCl, 90°C (● ): IC 50 = 25.1 ± 2.6 mM, n = 1.14 ± 0.13; tetracaine, 5 mM KCl, 90 °C (△): IC 50 = 4.2 ± 0.6 mM, n = 1.43 ± 0.33; tetracaine, 0 KCl, 22 °C (□ ): 1.2 ± 0.2 mM, n = 0.98 ± 0.22.
Figure Legend Snippet: Figure 2. Destabilization of KcsA tetramer as function of LA concentration. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing concentrations of lidocaine (●) or tetracaine (△, □ ). The sample assay mixture contained 10 mM Hepes-Tris, pH 7.5, 100 mM cholineCl and either 5 mM KCl (● , △) or no added KCl (□ ). Samples were incubated either at 90°C (● , △) or 22 °C (□ ) for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of LA concentration as described in Materials and Methods . Fit parameters: lidocaine, 5 mM KCl, 90°C (● ): IC 50 = 25.1 ± 2.6 mM, n = 1.14 ± 0.13; tetracaine, 5 mM KCl, 90 °C (△): IC 50 = 4.2 ± 0.6 mM, n = 1.43 ± 0.33; tetracaine, 0 KCl, 22 °C (□ ): 1.2 ± 0.2 mM, n = 0.98 ± 0.22.

Techniques Used: Concentration Assay, SDS Page, Incubation

Figure 3. Stabilization of KcsA tetramer as a function of K + concentration in the absence and presence of LA. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing K + concentration in the absence of LA (○) or in the presence of 20 mM lidocaine (● ) or 5 mM tetracaine (△). The sample assay mixture contained ~500 ng KcsA, 10 mM Hepes-Tris, pH 7.4, 100 mM choline Cl, indicated concentrations of KCl and either no LA, 20 mM lidocaine or 5 mM tetracaine. All samples were incubated at 90 °C for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of K + concentration as described in Materials and Methods . Fit parameters: no LA (○): K 0.5 = 1.48 ± 0.04 mM, n = 3.19 ± 0.26; 20 mM lidocaine (● ): K 0.5 = 6.65 ± 0.8 mM, n = 1.05 ± 0.13; 5 mM tetracaine (△): K 0.5 = 5.03 ± 0.90, n = 1.04 ± 0.19.
Figure Legend Snippet: Figure 3. Stabilization of KcsA tetramer as a function of K + concentration in the absence and presence of LA. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing K + concentration in the absence of LA (○) or in the presence of 20 mM lidocaine (● ) or 5 mM tetracaine (△). The sample assay mixture contained ~500 ng KcsA, 10 mM Hepes-Tris, pH 7.4, 100 mM choline Cl, indicated concentrations of KCl and either no LA, 20 mM lidocaine or 5 mM tetracaine. All samples were incubated at 90 °C for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of K + concentration as described in Materials and Methods . Fit parameters: no LA (○): K 0.5 = 1.48 ± 0.04 mM, n = 3.19 ± 0.26; 20 mM lidocaine (● ): K 0.5 = 6.65 ± 0.8 mM, n = 1.05 ± 0.13; 5 mM tetracaine (△): K 0.5 = 5.03 ± 0.90, n = 1.04 ± 0.19.

Techniques Used: Concentration Assay, SDS Page, Incubation

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    Bio-Rad acrylamide tris hcl ready sds page gels
    Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by <t>SDS-PAGE</t> analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM <t>Hepes-Tris,</t> pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.
    Acrylamide Tris Hcl Ready Sds Page Gels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by SDS-PAGE analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM Hepes-Tris, pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.

    Journal: Channels

    Article Title: Interaction of local anesthetics with the K+ channel pore domain

    doi: 10.4161/chan.24455

    Figure Lengend Snippet: Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by SDS-PAGE analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM Hepes-Tris, pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.

    Article Snippet: Ten milliliters of each sample was pipetted into wells of precast 12% acrylamide TRIS-HCl ready SDS-PAGE Gels (Bio-Rad) and subjected to electrophoresis at 200V for ~35 min in running buffer (25 mM Tris base, 192 mM glycine, pH 8.3, 0.1% SDS).

    Techniques: SDS Page, Incubation

    Figure 2. Destabilization of KcsA tetramer as function of LA concentration. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing concentrations of lidocaine (●) or tetracaine (△, □ ). The sample assay mixture contained 10 mM Hepes-Tris, pH 7.5, 100 mM cholineCl and either 5 mM KCl (● , △) or no added KCl (□ ). Samples were incubated either at 90°C (● , △) or 22 °C (□ ) for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of LA concentration as described in Materials and Methods . Fit parameters: lidocaine, 5 mM KCl, 90°C (● ): IC 50 = 25.1 ± 2.6 mM, n = 1.14 ± 0.13; tetracaine, 5 mM KCl, 90 °C (△): IC 50 = 4.2 ± 0.6 mM, n = 1.43 ± 0.33; tetracaine, 0 KCl, 22 °C (□ ): 1.2 ± 0.2 mM, n = 0.98 ± 0.22.

    Journal: Channels

    Article Title: Interaction of local anesthetics with the K+ channel pore domain

    doi: 10.4161/chan.24455

    Figure Lengend Snippet: Figure 2. Destabilization of KcsA tetramer as function of LA concentration. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing concentrations of lidocaine (●) or tetracaine (△, □ ). The sample assay mixture contained 10 mM Hepes-Tris, pH 7.5, 100 mM cholineCl and either 5 mM KCl (● , △) or no added KCl (□ ). Samples were incubated either at 90°C (● , △) or 22 °C (□ ) for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of LA concentration as described in Materials and Methods . Fit parameters: lidocaine, 5 mM KCl, 90°C (● ): IC 50 = 25.1 ± 2.6 mM, n = 1.14 ± 0.13; tetracaine, 5 mM KCl, 90 °C (△): IC 50 = 4.2 ± 0.6 mM, n = 1.43 ± 0.33; tetracaine, 0 KCl, 22 °C (□ ): 1.2 ± 0.2 mM, n = 0.98 ± 0.22.

    Article Snippet: Ten milliliters of each sample was pipetted into wells of precast 12% acrylamide TRIS-HCl ready SDS-PAGE Gels (Bio-Rad) and subjected to electrophoresis at 200V for ~35 min in running buffer (25 mM Tris base, 192 mM glycine, pH 8.3, 0.1% SDS).

    Techniques: Concentration Assay, SDS Page, Incubation

    Figure 3. Stabilization of KcsA tetramer as a function of K + concentration in the absence and presence of LA. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing K + concentration in the absence of LA (○) or in the presence of 20 mM lidocaine (● ) or 5 mM tetracaine (△). The sample assay mixture contained ~500 ng KcsA, 10 mM Hepes-Tris, pH 7.4, 100 mM choline Cl, indicated concentrations of KCl and either no LA, 20 mM lidocaine or 5 mM tetracaine. All samples were incubated at 90 °C for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of K + concentration as described in Materials and Methods . Fit parameters: no LA (○): K 0.5 = 1.48 ± 0.04 mM, n = 3.19 ± 0.26; 20 mM lidocaine (● ): K 0.5 = 6.65 ± 0.8 mM, n = 1.05 ± 0.13; 5 mM tetracaine (△): K 0.5 = 5.03 ± 0.90, n = 1.04 ± 0.19.

    Journal: Channels

    Article Title: Interaction of local anesthetics with the K+ channel pore domain

    doi: 10.4161/chan.24455

    Figure Lengend Snippet: Figure 3. Stabilization of KcsA tetramer as a function of K + concentration in the absence and presence of LA. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing K + concentration in the absence of LA (○) or in the presence of 20 mM lidocaine (● ) or 5 mM tetracaine (△). The sample assay mixture contained ~500 ng KcsA, 10 mM Hepes-Tris, pH 7.4, 100 mM choline Cl, indicated concentrations of KCl and either no LA, 20 mM lidocaine or 5 mM tetracaine. All samples were incubated at 90 °C for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of K + concentration as described in Materials and Methods . Fit parameters: no LA (○): K 0.5 = 1.48 ± 0.04 mM, n = 3.19 ± 0.26; 20 mM lidocaine (● ): K 0.5 = 6.65 ± 0.8 mM, n = 1.05 ± 0.13; 5 mM tetracaine (△): K 0.5 = 5.03 ± 0.90, n = 1.04 ± 0.19.

    Article Snippet: Ten milliliters of each sample was pipetted into wells of precast 12% acrylamide TRIS-HCl ready SDS-PAGE Gels (Bio-Rad) and subjected to electrophoresis at 200V for ~35 min in running buffer (25 mM Tris base, 192 mM glycine, pH 8.3, 0.1% SDS).

    Techniques: Concentration Assay, SDS Page, Incubation