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    Alomone Labs 1 ebio
    1 Ebio, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 1 article reviews
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    1 ebio - by Bioz Stars, 2022-01
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    Alomone Labs nimodipine
    Gonadotropin-releasing hormone (GnRH)-induced ERK1/2 phosphorylation is Gα q/11 and protein kinase C (PKC) dependent, and calcium independent in homologous LβT2 cells. ( A ) LβT2 cells were pretreated with 10 µM FR900359 (Gα q/11 inhibitor) or vehicle (dimethyl sulfoxide, DMSO) for 1 hr, and then treated with vehicle (water) or 10 nM GnRH for 5 or 15 min. Whole cell protein lysates were collected and subjected to sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blotting with phospho- (top) or total (bottom) ERK1/2 antibodies. Blots from one of two replicate experiments are shown. ( B ) Data from independent duplicate experiments exemplified in panel A were quantified by normalizing the densitometry for the pERK1/2 bands to the total ERK1/2 bands. Data are presented as fold phospho-ERK1/2 relative to the control condition. Bar height reflects the group mean. Data were analyzed by two-way analysis of variance (ANOVA), followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Vehicle vs. FR900359: 0 min GnRH p > 0.9999; 5 min GnRH p = 0.0106; 15 min GnRH p = 0.0085. ( C ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO) or 5 µM Gö6983 for 20 min followed by treatment with vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( D ) Data from three independent experiments in exemplified panel C were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control (vehicle) vs. GnRH (vehicle) p = 0.0048; Control (Gö6983) vs. GnRH (Gö6983) p = 0.3470. Control (vehicle) vs. Control (Gö6983) p = 0.9472; GnRH (vehicle) vs. GnRH (Gö6983) p = 0.0389. ( E ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO), 20 µM BAPTA-AM, 5 µM or 10 µM <t>nimodipine,</t> or 10 µM U0126 for 20 min followed by vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( F ) Data from three independent experiments in panel E were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control vs. GnRH: Vehicle p = 0.0103; BAPTA-AM p = 0.0229; Nimodipine (5 µM) p = 0.0377; Nimodipine (10 µM) p = 0.0350; U1026 p > 0.9999. Source data for Figure 7—figure supplement 1A . Source data for Figure 7—figure supplement 1C . Source data for Figure 7—figure supplement 1E .
    Nimodipine, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs 1 ebio
    Gonadotropin-releasing hormone (GnRH)-induced ERK1/2 phosphorylation is Gα q/11 and protein kinase C (PKC) dependent, and calcium independent in homologous LβT2 cells. ( A ) LβT2 cells were pretreated with 10 µM FR900359 (Gα q/11 inhibitor) or vehicle (dimethyl sulfoxide, DMSO) for 1 hr, and then treated with vehicle (water) or 10 nM GnRH for 5 or 15 min. Whole cell protein lysates were collected and subjected to sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blotting with phospho- (top) or total (bottom) ERK1/2 antibodies. Blots from one of two replicate experiments are shown. ( B ) Data from independent duplicate experiments exemplified in panel A were quantified by normalizing the densitometry for the pERK1/2 bands to the total ERK1/2 bands. Data are presented as fold phospho-ERK1/2 relative to the control condition. Bar height reflects the group mean. Data were analyzed by two-way analysis of variance (ANOVA), followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Vehicle vs. FR900359: 0 min GnRH p > 0.9999; 5 min GnRH p = 0.0106; 15 min GnRH p = 0.0085. ( C ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO) or 5 µM Gö6983 for 20 min followed by treatment with vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( D ) Data from three independent experiments in exemplified panel C were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control (vehicle) vs. GnRH (vehicle) p = 0.0048; Control (Gö6983) vs. GnRH (Gö6983) p = 0.3470. Control (vehicle) vs. Control (Gö6983) p = 0.9472; GnRH (vehicle) vs. GnRH (Gö6983) p = 0.0389. ( E ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO), 20 µM BAPTA-AM, 5 µM or 10 µM <t>nimodipine,</t> or 10 µM U0126 for 20 min followed by vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( F ) Data from three independent experiments in panel E were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control vs. GnRH: Vehicle p = 0.0103; BAPTA-AM p = 0.0229; Nimodipine (5 µM) p = 0.0377; Nimodipine (10 µM) p = 0.0350; U1026 p > 0.9999. Source data for Figure 7—figure supplement 1A . Source data for Figure 7—figure supplement 1C . Source data for Figure 7—figure supplement 1E .
    1 Ebio, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gonadotropin-releasing hormone (GnRH)-induced ERK1/2 phosphorylation is Gα q/11 and protein kinase C (PKC) dependent, and calcium independent in homologous LβT2 cells. ( A ) LβT2 cells were pretreated with 10 µM FR900359 (Gα q/11 inhibitor) or vehicle (dimethyl sulfoxide, DMSO) for 1 hr, and then treated with vehicle (water) or 10 nM GnRH for 5 or 15 min. Whole cell protein lysates were collected and subjected to sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blotting with phospho- (top) or total (bottom) ERK1/2 antibodies. Blots from one of two replicate experiments are shown. ( B ) Data from independent duplicate experiments exemplified in panel A were quantified by normalizing the densitometry for the pERK1/2 bands to the total ERK1/2 bands. Data are presented as fold phospho-ERK1/2 relative to the control condition. Bar height reflects the group mean. Data were analyzed by two-way analysis of variance (ANOVA), followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Vehicle vs. FR900359: 0 min GnRH p > 0.9999; 5 min GnRH p = 0.0106; 15 min GnRH p = 0.0085. ( C ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO) or 5 µM Gö6983 for 20 min followed by treatment with vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( D ) Data from three independent experiments in exemplified panel C were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control (vehicle) vs. GnRH (vehicle) p = 0.0048; Control (Gö6983) vs. GnRH (Gö6983) p = 0.3470. Control (vehicle) vs. Control (Gö6983) p = 0.9472; GnRH (vehicle) vs. GnRH (Gö6983) p = 0.0389. ( E ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO), 20 µM BAPTA-AM, 5 µM or 10 µM nimodipine, or 10 µM U0126 for 20 min followed by vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( F ) Data from three independent experiments in panel E were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control vs. GnRH: Vehicle p = 0.0103; BAPTA-AM p = 0.0229; Nimodipine (5 µM) p = 0.0377; Nimodipine (10 µM) p = 0.0350; U1026 p > 0.9999. Source data for Figure 7—figure supplement 1A . Source data for Figure 7—figure supplement 1C . Source data for Figure 7—figure supplement 1E .

    Journal: eLife

    Article Title: Addition of a carboxy-terminal tail to the normally tailless gonadotropin-releasing hormone receptor impairs fertility in female mice

    doi: 10.7554/eLife.72937

    Figure Lengend Snippet: Gonadotropin-releasing hormone (GnRH)-induced ERK1/2 phosphorylation is Gα q/11 and protein kinase C (PKC) dependent, and calcium independent in homologous LβT2 cells. ( A ) LβT2 cells were pretreated with 10 µM FR900359 (Gα q/11 inhibitor) or vehicle (dimethyl sulfoxide, DMSO) for 1 hr, and then treated with vehicle (water) or 10 nM GnRH for 5 or 15 min. Whole cell protein lysates were collected and subjected to sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blotting with phospho- (top) or total (bottom) ERK1/2 antibodies. Blots from one of two replicate experiments are shown. ( B ) Data from independent duplicate experiments exemplified in panel A were quantified by normalizing the densitometry for the pERK1/2 bands to the total ERK1/2 bands. Data are presented as fold phospho-ERK1/2 relative to the control condition. Bar height reflects the group mean. Data were analyzed by two-way analysis of variance (ANOVA), followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Vehicle vs. FR900359: 0 min GnRH p > 0.9999; 5 min GnRH p = 0.0106; 15 min GnRH p = 0.0085. ( C ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO) or 5 µM Gö6983 for 20 min followed by treatment with vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( D ) Data from three independent experiments in exemplified panel C were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control (vehicle) vs. GnRH (vehicle) p = 0.0048; Control (Gö6983) vs. GnRH (Gö6983) p = 0.3470. Control (vehicle) vs. Control (Gö6983) p = 0.9472; GnRH (vehicle) vs. GnRH (Gö6983) p = 0.0389. ( E ) LβT2 cells were pretreated for 20 min with vehicle (dimethyl sulfoxide, DMSO), 20 µM BAPTA-AM, 5 µM or 10 µM nimodipine, or 10 µM U0126 for 20 min followed by vehicle (water) or 10 nM GnRH for 5 min. Western blots were performed as in panel A. ( F ) Data from three independent experiments in panel E were presented and quantified as in panel B. Data were analyzed by two-way ANOVA, followed by Sidak’s post hoc comparison tests. Bars with different letters differed significantly. Control vs. GnRH: Vehicle p = 0.0103; BAPTA-AM p = 0.0229; Nimodipine (5 µM) p = 0.0377; Nimodipine (10 µM) p = 0.0350; U1026 p > 0.9999. Source data for Figure 7—figure supplement 1A . Source data for Figure 7—figure supplement 1C . Source data for Figure 7—figure supplement 1E .

    Article Snippet: To evaluate the contribution of voltage-gated calcium channels, after 1 hr of recovery, tissue was incubated for 30 s with 20 μM nimodipine (ALOMONE LABS N-150 N150SM0250; Jerusalem, Israel) followed by a second application of 10 nM GnRH alone ( ) or in combination with 20 μM nimodipine ( ) for 30 s. Finally, to determine cell viability, high potassium solution (50 KCl mM, 120 NaCl mM, 10 HEPES mM, 2 CaCl2 mM, pH 7.4) was applied for 30 s. For each condition, the numbers of animals and cells analyzed are indicated in the figure legends.

    Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot

    Gonadotropin-releasing hormone (GnRH)-induced Fshb and Lhb expression does not depend on calcium entry via L-type channels in homologous LβT2 cells. ( A ) Relative Fshb and ( B ) Lhb expression in LβT2 cells treated with vehicle (dimethyl sulfoxide, DMSO) or 10 µM nimodipine for 20 min followed by treatment with water (vehicle) or high-frequency GnRH (10 nM) pulses. Gene expression was assessed by RT-qPCR and normalized to Rpl19 . Data reflect the means of two independent experiments. Data were analyzed with two-way analyses of variance (ANOVAs), followed by post hoc Tukey’s test for multiple comparisons. Bars with different letters differed significantly. Panel A: Control (vehicle) vs. GnRH (vehicle) p = 0.0178; Control (vehicle) vs. GnRH (Nimodipine) p = 0.0232; GnRH (vehicle) vs. GnRH (Nimodipine) p = 0.8545; Control (Nimodipine) vs. GnRH (Nimodipine) p = 0.0235. Panel B: Control (vehicle) vs. GnRH (vehicle) p = 0.0061; Control (vehicle) vs. GnRH (Nimodipine) p = 0.0043; GnRH (vehicle) vs. GnRH (Nimodipine) p = 0.8770; Control (Nimodipine) vs. GnRH (Nimodipine) p = 0.0178.

    Journal: eLife

    Article Title: Addition of a carboxy-terminal tail to the normally tailless gonadotropin-releasing hormone receptor impairs fertility in female mice

    doi: 10.7554/eLife.72937

    Figure Lengend Snippet: Gonadotropin-releasing hormone (GnRH)-induced Fshb and Lhb expression does not depend on calcium entry via L-type channels in homologous LβT2 cells. ( A ) Relative Fshb and ( B ) Lhb expression in LβT2 cells treated with vehicle (dimethyl sulfoxide, DMSO) or 10 µM nimodipine for 20 min followed by treatment with water (vehicle) or high-frequency GnRH (10 nM) pulses. Gene expression was assessed by RT-qPCR and normalized to Rpl19 . Data reflect the means of two independent experiments. Data were analyzed with two-way analyses of variance (ANOVAs), followed by post hoc Tukey’s test for multiple comparisons. Bars with different letters differed significantly. Panel A: Control (vehicle) vs. GnRH (vehicle) p = 0.0178; Control (vehicle) vs. GnRH (Nimodipine) p = 0.0232; GnRH (vehicle) vs. GnRH (Nimodipine) p = 0.8545; Control (Nimodipine) vs. GnRH (Nimodipine) p = 0.0235. Panel B: Control (vehicle) vs. GnRH (vehicle) p = 0.0061; Control (vehicle) vs. GnRH (Nimodipine) p = 0.0043; GnRH (vehicle) vs. GnRH (Nimodipine) p = 0.8770; Control (Nimodipine) vs. GnRH (Nimodipine) p = 0.0178.

    Article Snippet: To evaluate the contribution of voltage-gated calcium channels, after 1 hr of recovery, tissue was incubated for 30 s with 20 μM nimodipine (ALOMONE LABS N-150 N150SM0250; Jerusalem, Israel) followed by a second application of 10 nM GnRH alone ( ) or in combination with 20 μM nimodipine ( ) for 30 s. Finally, to determine cell viability, high potassium solution (50 KCl mM, 120 NaCl mM, 10 HEPES mM, 2 CaCl2 mM, pH 7.4) was applied for 30 s. For each condition, the numbers of animals and cells analyzed are indicated in the figure legends.

    Techniques: Expressing, Quantitative RT-PCR

    Nimodipine alters gonadotropin-releasing hormone (GnRH)-induced calcium responses in gonadotropes of wild-type and Ctail mice. The analysis in Figure 8—figure supplement 1 was repeated but with the L-type calcium channel blocker nimodipine (Nim) applied prior to and during the second GnRH pulse. Raster plots of GnRH-induced calcium responses in the absence (left) and presence of nimodipine (right) in gonadotropes from a representative adult male ( A ) wild-type and ( D ) Ctail mouse. The two stimuli were separated by 60 min wash with Krebs–Ringer. Comparisons of AUC from a ( B ) wild-type (275 cells; 11.5 ± 4.7 vs. 7.1 ± 2.8 a.u. for GnRH and GnRH/Nim, respectively; p

    Journal: eLife

    Article Title: Addition of a carboxy-terminal tail to the normally tailless gonadotropin-releasing hormone receptor impairs fertility in female mice

    doi: 10.7554/eLife.72937

    Figure Lengend Snippet: Nimodipine alters gonadotropin-releasing hormone (GnRH)-induced calcium responses in gonadotropes of wild-type and Ctail mice. The analysis in Figure 8—figure supplement 1 was repeated but with the L-type calcium channel blocker nimodipine (Nim) applied prior to and during the second GnRH pulse. Raster plots of GnRH-induced calcium responses in the absence (left) and presence of nimodipine (right) in gonadotropes from a representative adult male ( A ) wild-type and ( D ) Ctail mouse. The two stimuli were separated by 60 min wash with Krebs–Ringer. Comparisons of AUC from a ( B ) wild-type (275 cells; 11.5 ± 4.7 vs. 7.1 ± 2.8 a.u. for GnRH and GnRH/Nim, respectively; p

    Article Snippet: To evaluate the contribution of voltage-gated calcium channels, after 1 hr of recovery, tissue was incubated for 30 s with 20 μM nimodipine (ALOMONE LABS N-150 N150SM0250; Jerusalem, Israel) followed by a second application of 10 nM GnRH alone ( ) or in combination with 20 μM nimodipine ( ) for 30 s. Finally, to determine cell viability, high potassium solution (50 KCl mM, 120 NaCl mM, 10 HEPES mM, 2 CaCl2 mM, pH 7.4) was applied for 30 s. For each condition, the numbers of animals and cells analyzed are indicated in the figure legends.

    Techniques: Mouse Assay