rabbit anti kv7 1 antibody  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti kv7 1 antibody
    Mutation of phosphorylated serines does not affect subcellular localization of <t>Kv7.1.</t> Alanine ( A ) and aspartate ( B ) mutants of the identified serine phosphorylation sites were coexpressed with the ER marker pDsRed2-ER in MDCK cells. The cells were stained
    Rabbit Anti Kv7 1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti kv7 1 antibody/product/Alomone Labs
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rabbit anti kv7 1 antibody - by Bioz Stars, 2022-08
    90/100 stars

    Images

    1) Product Images from "Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking"

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00383.2014

    Mutation of phosphorylated serines does not affect subcellular localization of Kv7.1. Alanine ( A ) and aspartate ( B ) mutants of the identified serine phosphorylation sites were coexpressed with the ER marker pDsRed2-ER in MDCK cells. The cells were stained
    Figure Legend Snippet: Mutation of phosphorylated serines does not affect subcellular localization of Kv7.1. Alanine ( A ) and aspartate ( B ) mutants of the identified serine phosphorylation sites were coexpressed with the ER marker pDsRed2-ER in MDCK cells. The cells were stained

    Techniques Used: Mutagenesis, Marker, Staining

    Summary of proteomics results for Kv7.1. A : topological representation of the Kv7.1 channel, including the amino acid sequence. The amino acid sequences highlighted in blue were covered in the mass spectrometry experiments, and the identified phosphorylation
    Figure Legend Snippet: Summary of proteomics results for Kv7.1. A : topological representation of the Kv7.1 channel, including the amino acid sequence. The amino acid sequences highlighted in blue were covered in the mass spectrometry experiments, and the identified phosphorylation

    Techniques Used: Sequencing, Mass Spectrometry

    Inhibition of PKA leads to internalization and possibly lysosomal degradation of Kv7.1.
    Figure Legend Snippet: Inhibition of PKA leads to internalization and possibly lysosomal degradation of Kv7.1.

    Techniques Used: Inhibition

    Kv7.1 accumulates intracellularly upon exposure to PKA inhibitors KT-5720 and myristoylated protein kinase A inhibitor 14–22 amide. A : MDCK cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h (control) followed by treatment
    Figure Legend Snippet: Kv7.1 accumulates intracellularly upon exposure to PKA inhibitors KT-5720 and myristoylated protein kinase A inhibitor 14–22 amide. A : MDCK cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h (control) followed by treatment

    Techniques Used: Stable Transfection, Expressing

    Kv7.1 codistributes with late endosomal/lysosomal markers in response to H-89 treatment. A : confocal scans of MDCK-Kv7.1 cells treated with 20 μM H-89 for 3 h and colabeled with the late endosomal/lysosomal marker LAMP-2. Scale bar = 10 ( top )
    Figure Legend Snippet: Kv7.1 codistributes with late endosomal/lysosomal markers in response to H-89 treatment. A : confocal scans of MDCK-Kv7.1 cells treated with 20 μM H-89 for 3 h and colabeled with the late endosomal/lysosomal marker LAMP-2. Scale bar = 10 ( top )

    Techniques Used: Marker

    Small-interfering RNA (siRNA) knockdown of Nedd4-2 protects Kv7.1 from internalization upon PKA inhibition. A : MDCK cells stably expressing Kv7.1 were transiently transfected with siRNA targeting Nedd4-2 or with noncoding control siRNA. Enhanced GFP (eGFP)
    Figure Legend Snippet: Small-interfering RNA (siRNA) knockdown of Nedd4-2 protects Kv7.1 from internalization upon PKA inhibition. A : MDCK cells stably expressing Kv7.1 were transiently transfected with siRNA targeting Nedd4-2 or with noncoding control siRNA. Enhanced GFP (eGFP)

    Techniques Used: Small Interfering RNA, Inhibition, Stable Transfection, Expressing, Transfection

    Kv7.1-YA localization is unaffected by PKA inhibition. MDCK cells stably expressing the Kv7.1-YA mutant were treated with 20 μM H-89 for 3–5 h. A : confocal scans of MDCK-Kv7.1-YA cells before (0 h) and 3 (3 h H-89) and 5 (5 h H-89) h after
    Figure Legend Snippet: Kv7.1-YA localization is unaffected by PKA inhibition. MDCK cells stably expressing the Kv7.1-YA mutant were treated with 20 μM H-89 for 3–5 h. A : confocal scans of MDCK-Kv7.1-YA cells before (0 h) and 3 (3 h H-89) and 5 (5 h H-89) h after

    Techniques Used: Inhibition, Stable Transfection, Expressing, Mutagenesis

    Kv7.1 redistribution is not caused by increased Nedd4-2 binding or ubiquitinylation. A : representative Western blot showing the effects of 20 μM H-89 (3 h exposure) and 50 μM forskolin (1 h exposure) on the abundance of S 468 -phosphorylated
    Figure Legend Snippet: Kv7.1 redistribution is not caused by increased Nedd4-2 binding or ubiquitinylation. A : representative Western blot showing the effects of 20 μM H-89 (3 h exposure) and 50 μM forskolin (1 h exposure) on the abundance of S 468 -phosphorylated

    Techniques Used: Binding Assay, Western Blot

    Kv7.1 accumulates intracellularly upon inhibition of protein kinase A (PKA). Madin-Darby canine kidney (MDCK) cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h followed by treatment with 20 μM H-89 for 3–5 h. A
    Figure Legend Snippet: Kv7.1 accumulates intracellularly upon inhibition of protein kinase A (PKA). Madin-Darby canine kidney (MDCK) cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h followed by treatment with 20 μM H-89 for 3–5 h. A

    Techniques Used: Inhibition, Stable Transfection, Expressing

    Intracellular accumulation in response to PKA inhibition is specific for Kv7.1. MDCK cells were transfected with Kv7.4 and allowed to polarize before being subjected to either 20 μM H-89 or 10 μM KT-5720 treatments for 3 h. As demonstrated,
    Figure Legend Snippet: Intracellular accumulation in response to PKA inhibition is specific for Kv7.1. MDCK cells were transfected with Kv7.4 and allowed to polarize before being subjected to either 20 μM H-89 or 10 μM KT-5720 treatments for 3 h. As demonstrated,

    Techniques Used: Inhibition, Transfection

    PKA activation promotes Kv7.1 surface expression in polarized MDCK cells. MDCK cells stably expressing Kv7.1 were grown to confluence and allowed to polarize. The cells were then treated with 50 μM forskolin for 1 h. Cells were fixed before and
    Figure Legend Snippet: PKA activation promotes Kv7.1 surface expression in polarized MDCK cells. MDCK cells stably expressing Kv7.1 were grown to confluence and allowed to polarize. The cells were then treated with 50 μM forskolin for 1 h. Cells were fixed before and

    Techniques Used: Activation Assay, Expressing, Stable Transfection

    Mimicked phosphorylation of T470 results in reduced membrane expression of Kv7.1, but Kv7.1-T470A is still endocytosed in response to PKA inhibition. Kv7.1-T470A and Kv7.1-T470D mutants were coexpressed with DsRed2-ER in MDCK cells, which were allowed
    Figure Legend Snippet: Mimicked phosphorylation of T470 results in reduced membrane expression of Kv7.1, but Kv7.1-T470A is still endocytosed in response to PKA inhibition. Kv7.1-T470A and Kv7.1-T470D mutants were coexpressed with DsRed2-ER in MDCK cells, which were allowed

    Techniques Used: Expressing, Inhibition

    2) Product Images from "Inactivation gating of Kv7.1 channels does not involve concerted cooperative subunit interactions"

    Article Title: Inactivation gating of Kv7.1 channels does not involve concerted cooperative subunit interactions

    Journal: Channels

    doi: 10.1080/19336950.2018.1441649

    Expression of concatenated Kv7.1 subunits bearing an increasing number of L273F mutations. A, representative current traces of the following constructs: ConD 2 L273F (ConLFLL), ConD 2,4 L273F (ConLFLF), ConD 1,2 L273F (ConFFLL), ConD ,2,3,4 L273F (ConLFFF) and ConD 1,2,3,4 L273F (ConFFFF). CHO cells were held at −90 mV and the membrane voltage was stepped for 2 s from −70 mV to +50 mV in 10 mV increments and then repolarized for 1 s to −60 mV. B, Fractional inactivation F of the different concatemer constructs was measured at +50 mV (n = 7–10). C, Time constants of inactivation relaxation measured at +50 mV and determined by one exponential fit of the inactivation traces (n = 8–14). D, conductance-voltage relations of the different concatemer constructs. Data were fitted with a single Boltzmann function (n = 7–10).
    Figure Legend Snippet: Expression of concatenated Kv7.1 subunits bearing an increasing number of L273F mutations. A, representative current traces of the following constructs: ConD 2 L273F (ConLFLL), ConD 2,4 L273F (ConLFLF), ConD 1,2 L273F (ConFFLL), ConD ,2,3,4 L273F (ConLFFF) and ConD 1,2,3,4 L273F (ConFFFF). CHO cells were held at −90 mV and the membrane voltage was stepped for 2 s from −70 mV to +50 mV in 10 mV increments and then repolarized for 1 s to −60 mV. B, Fractional inactivation F of the different concatemer constructs was measured at +50 mV (n = 7–10). C, Time constants of inactivation relaxation measured at +50 mV and determined by one exponential fit of the inactivation traces (n = 8–14). D, conductance-voltage relations of the different concatemer constructs. Data were fitted with a single Boltzmann function (n = 7–10).

    Techniques Used: Expressing, Construct

    Characterization of the WT and L273F mutant Kv7.1 concatenated tetrameric channels. A, Top panel, scheme of the Kv7.1 concatenated tetrameric channel construct (Con), where subunits D1, D2, D3 and D4 are connected by flexible linkers. Each linker (8 glycines) harbors a unique restriction site. B, conductance-voltage relations of WT monomeric and WT concatenated tetrameric Kv7.1 constructs in the absence or presence of KCNE1. Data were fitted with a single Boltzmann function. C, Western blot showing lysates from HEK293 transfected with empty vector (Mock), KCNQ1 monomeric and concatenated tetrameric constructs. D, location of residue L273 in the S5 α helical segment of the Kv7.1 pore domain as mapped in the recent cryo-EM structure of the frog Kv7.1 (Sun, 2017). E, representative current traces of L273F Kv7.1 monomeric and concatenated tetrameric mutant constructs. CHO cells were held at −90 mV and the membrane voltage was stepped for 3 s from −60 mV to +60 mV in 10 mV increments and then repolarized for 1.5 s to −60 mV. F, conductance-voltage relations of the WT Kv7.1 concatemer, L273F Kv7.1 monomer and L273F Kv7.1 concatemer. Data were fitted with a single Boltzmann function.
    Figure Legend Snippet: Characterization of the WT and L273F mutant Kv7.1 concatenated tetrameric channels. A, Top panel, scheme of the Kv7.1 concatenated tetrameric channel construct (Con), where subunits D1, D2, D3 and D4 are connected by flexible linkers. Each linker (8 glycines) harbors a unique restriction site. B, conductance-voltage relations of WT monomeric and WT concatenated tetrameric Kv7.1 constructs in the absence or presence of KCNE1. Data were fitted with a single Boltzmann function. C, Western blot showing lysates from HEK293 transfected with empty vector (Mock), KCNQ1 monomeric and concatenated tetrameric constructs. D, location of residue L273 in the S5 α helical segment of the Kv7.1 pore domain as mapped in the recent cryo-EM structure of the frog Kv7.1 (Sun, 2017). E, representative current traces of L273F Kv7.1 monomeric and concatenated tetrameric mutant constructs. CHO cells were held at −90 mV and the membrane voltage was stepped for 3 s from −60 mV to +60 mV in 10 mV increments and then repolarized for 1.5 s to −60 mV. F, conductance-voltage relations of the WT Kv7.1 concatemer, L273F Kv7.1 monomer and L273F Kv7.1 concatemer. Data were fitted with a single Boltzmann function.

    Techniques Used: Mutagenesis, Construct, Western Blot, Transfection, Plasmid Preparation, Cryo-EM Sample Prep

    Recovery from inactivation of the different concatemer constructs. A, representative current traces of the recovery from inactivation of the different concatemer constructs. The recovery time from inactivation was determined from a −90 mV holding potential, by stepping the membrane voltage for 1.5 s to +50 mV to open the various Kv7.1 concatemer channels (first pulse), then repolarizing to −90 mV for various times to allow recovery from inactivation and stepping back to +50 mV for 0.5 s to reopen the channels (second pulse). B, The fractional recovery was calculated by the peak current of the second pulse divided by the peak of the first pulse. These ratios were plotted against the recovery times (n = 4). C, time constants of the recovery time from inactivation were calculated from an exponential fit (one way ANOVA, n = 4; * P
    Figure Legend Snippet: Recovery from inactivation of the different concatemer constructs. A, representative current traces of the recovery from inactivation of the different concatemer constructs. The recovery time from inactivation was determined from a −90 mV holding potential, by stepping the membrane voltage for 1.5 s to +50 mV to open the various Kv7.1 concatemer channels (first pulse), then repolarizing to −90 mV for various times to allow recovery from inactivation and stepping back to +50 mV for 0.5 s to reopen the channels (second pulse). B, The fractional recovery was calculated by the peak current of the second pulse divided by the peak of the first pulse. These ratios were plotted against the recovery times (n = 4). C, time constants of the recovery time from inactivation were calculated from an exponential fit (one way ANOVA, n = 4; * P

    Techniques Used: Construct

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    Alomone Labs rabbit anti kv7 1 antibody
    Mutation of phosphorylated serines does not affect subcellular localization of <t>Kv7.1.</t> Alanine ( A ) and aspartate ( B ) mutants of the identified serine phosphorylation sites were coexpressed with the ER marker pDsRed2-ER in MDCK cells. The cells were stained
    Rabbit Anti Kv7 1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti kv7 1 antibody/product/Alomone Labs
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti kv7 1 antibody - by Bioz Stars, 2022-08
    90/100 stars
      Buy from Supplier

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    Mutation of phosphorylated serines does not affect subcellular localization of Kv7.1. Alanine ( A ) and aspartate ( B ) mutants of the identified serine phosphorylation sites were coexpressed with the ER marker pDsRed2-ER in MDCK cells. The cells were stained

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Mutation of phosphorylated serines does not affect subcellular localization of Kv7.1. Alanine ( A ) and aspartate ( B ) mutants of the identified serine phosphorylation sites were coexpressed with the ER marker pDsRed2-ER in MDCK cells. The cells were stained

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Mutagenesis, Marker, Staining

    Summary of proteomics results for Kv7.1. A : topological representation of the Kv7.1 channel, including the amino acid sequence. The amino acid sequences highlighted in blue were covered in the mass spectrometry experiments, and the identified phosphorylation

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Summary of proteomics results for Kv7.1. A : topological representation of the Kv7.1 channel, including the amino acid sequence. The amino acid sequences highlighted in blue were covered in the mass spectrometry experiments, and the identified phosphorylation

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Sequencing, Mass Spectrometry

    Inhibition of PKA leads to internalization and possibly lysosomal degradation of Kv7.1.

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Inhibition of PKA leads to internalization and possibly lysosomal degradation of Kv7.1.

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Inhibition

    Kv7.1 accumulates intracellularly upon exposure to PKA inhibitors KT-5720 and myristoylated protein kinase A inhibitor 14–22 amide. A : MDCK cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h (control) followed by treatment

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Kv7.1 accumulates intracellularly upon exposure to PKA inhibitors KT-5720 and myristoylated protein kinase A inhibitor 14–22 amide. A : MDCK cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h (control) followed by treatment

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Stable Transfection, Expressing

    Kv7.1 codistributes with late endosomal/lysosomal markers in response to H-89 treatment. A : confocal scans of MDCK-Kv7.1 cells treated with 20 μM H-89 for 3 h and colabeled with the late endosomal/lysosomal marker LAMP-2. Scale bar = 10 ( top )

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Kv7.1 codistributes with late endosomal/lysosomal markers in response to H-89 treatment. A : confocal scans of MDCK-Kv7.1 cells treated with 20 μM H-89 for 3 h and colabeled with the late endosomal/lysosomal marker LAMP-2. Scale bar = 10 ( top )

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Marker

    Small-interfering RNA (siRNA) knockdown of Nedd4-2 protects Kv7.1 from internalization upon PKA inhibition. A : MDCK cells stably expressing Kv7.1 were transiently transfected with siRNA targeting Nedd4-2 or with noncoding control siRNA. Enhanced GFP (eGFP)

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Small-interfering RNA (siRNA) knockdown of Nedd4-2 protects Kv7.1 from internalization upon PKA inhibition. A : MDCK cells stably expressing Kv7.1 were transiently transfected with siRNA targeting Nedd4-2 or with noncoding control siRNA. Enhanced GFP (eGFP)

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Small Interfering RNA, Inhibition, Stable Transfection, Expressing, Transfection

    Kv7.1-YA localization is unaffected by PKA inhibition. MDCK cells stably expressing the Kv7.1-YA mutant were treated with 20 μM H-89 for 3–5 h. A : confocal scans of MDCK-Kv7.1-YA cells before (0 h) and 3 (3 h H-89) and 5 (5 h H-89) h after

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Kv7.1-YA localization is unaffected by PKA inhibition. MDCK cells stably expressing the Kv7.1-YA mutant were treated with 20 μM H-89 for 3–5 h. A : confocal scans of MDCK-Kv7.1-YA cells before (0 h) and 3 (3 h H-89) and 5 (5 h H-89) h after

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Inhibition, Stable Transfection, Expressing, Mutagenesis

    Kv7.1 redistribution is not caused by increased Nedd4-2 binding or ubiquitinylation. A : representative Western blot showing the effects of 20 μM H-89 (3 h exposure) and 50 μM forskolin (1 h exposure) on the abundance of S 468 -phosphorylated

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Kv7.1 redistribution is not caused by increased Nedd4-2 binding or ubiquitinylation. A : representative Western blot showing the effects of 20 μM H-89 (3 h exposure) and 50 μM forskolin (1 h exposure) on the abundance of S 468 -phosphorylated

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Binding Assay, Western Blot

    Kv7.1 accumulates intracellularly upon inhibition of protein kinase A (PKA). Madin-Darby canine kidney (MDCK) cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h followed by treatment with 20 μM H-89 for 3–5 h. A

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Kv7.1 accumulates intracellularly upon inhibition of protein kinase A (PKA). Madin-Darby canine kidney (MDCK) cells stably expressing Kv7.1 were subjected to a calcium switch for 24 h followed by treatment with 20 μM H-89 for 3–5 h. A

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Inhibition, Stable Transfection, Expressing

    Intracellular accumulation in response to PKA inhibition is specific for Kv7.1. MDCK cells were transfected with Kv7.4 and allowed to polarize before being subjected to either 20 μM H-89 or 10 μM KT-5720 treatments for 3 h. As demonstrated,

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Intracellular accumulation in response to PKA inhibition is specific for Kv7.1. MDCK cells were transfected with Kv7.4 and allowed to polarize before being subjected to either 20 μM H-89 or 10 μM KT-5720 treatments for 3 h. As demonstrated,

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Inhibition, Transfection

    PKA activation promotes Kv7.1 surface expression in polarized MDCK cells. MDCK cells stably expressing Kv7.1 were grown to confluence and allowed to polarize. The cells were then treated with 50 μM forskolin for 1 h. Cells were fixed before and

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: PKA activation promotes Kv7.1 surface expression in polarized MDCK cells. MDCK cells stably expressing Kv7.1 were grown to confluence and allowed to polarize. The cells were then treated with 50 μM forskolin for 1 h. Cells were fixed before and

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Activation Assay, Expressing, Stable Transfection

    Mimicked phosphorylation of T470 results in reduced membrane expression of Kv7.1, but Kv7.1-T470A is still endocytosed in response to PKA inhibition. Kv7.1-T470A and Kv7.1-T470D mutants were coexpressed with DsRed2-ER in MDCK cells, which were allowed

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

    doi: 10.1152/ajpcell.00383.2014

    Figure Lengend Snippet: Mimicked phosphorylation of T470 results in reduced membrane expression of Kv7.1, but Kv7.1-T470A is still endocytosed in response to PKA inhibition. Kv7.1-T470A and Kv7.1-T470D mutants were coexpressed with DsRed2-ER in MDCK cells, which were allowed

    Article Snippet: The precleared lysate was subsequently incubated with 3 μg of rabbit anti-Kv7.1 antibody (Alomone Labs), or, as a control, 3 μg normal rabbit IgG (Santa Cruz Biotechnology) for 1 h at 4°C.

    Techniques: Expressing, Inhibition

    Expression of concatenated Kv7.1 subunits bearing an increasing number of L273F mutations. A, representative current traces of the following constructs: ConD 2 L273F (ConLFLL), ConD 2,4 L273F (ConLFLF), ConD 1,2 L273F (ConFFLL), ConD ,2,3,4 L273F (ConLFFF) and ConD 1,2,3,4 L273F (ConFFFF). CHO cells were held at −90 mV and the membrane voltage was stepped for 2 s from −70 mV to +50 mV in 10 mV increments and then repolarized for 1 s to −60 mV. B, Fractional inactivation F of the different concatemer constructs was measured at +50 mV (n = 7–10). C, Time constants of inactivation relaxation measured at +50 mV and determined by one exponential fit of the inactivation traces (n = 8–14). D, conductance-voltage relations of the different concatemer constructs. Data were fitted with a single Boltzmann function (n = 7–10).

    Journal: Channels

    Article Title: Inactivation gating of Kv7.1 channels does not involve concerted cooperative subunit interactions

    doi: 10.1080/19336950.2018.1441649

    Figure Lengend Snippet: Expression of concatenated Kv7.1 subunits bearing an increasing number of L273F mutations. A, representative current traces of the following constructs: ConD 2 L273F (ConLFLL), ConD 2,4 L273F (ConLFLF), ConD 1,2 L273F (ConFFLL), ConD ,2,3,4 L273F (ConLFFF) and ConD 1,2,3,4 L273F (ConFFFF). CHO cells were held at −90 mV and the membrane voltage was stepped for 2 s from −70 mV to +50 mV in 10 mV increments and then repolarized for 1 s to −60 mV. B, Fractional inactivation F of the different concatemer constructs was measured at +50 mV (n = 7–10). C, Time constants of inactivation relaxation measured at +50 mV and determined by one exponential fit of the inactivation traces (n = 8–14). D, conductance-voltage relations of the different concatemer constructs. Data were fitted with a single Boltzmann function (n = 7–10).

    Article Snippet: Equal amounts of lysate proteins were resolved by 8% SDS-PAGE and blots were reacted using rabbit anti-Kv7.1 antibodies (Alomone Labs).

    Techniques: Expressing, Construct

    Characterization of the WT and L273F mutant Kv7.1 concatenated tetrameric channels. A, Top panel, scheme of the Kv7.1 concatenated tetrameric channel construct (Con), where subunits D1, D2, D3 and D4 are connected by flexible linkers. Each linker (8 glycines) harbors a unique restriction site. B, conductance-voltage relations of WT monomeric and WT concatenated tetrameric Kv7.1 constructs in the absence or presence of KCNE1. Data were fitted with a single Boltzmann function. C, Western blot showing lysates from HEK293 transfected with empty vector (Mock), KCNQ1 monomeric and concatenated tetrameric constructs. D, location of residue L273 in the S5 α helical segment of the Kv7.1 pore domain as mapped in the recent cryo-EM structure of the frog Kv7.1 (Sun, 2017). E, representative current traces of L273F Kv7.1 monomeric and concatenated tetrameric mutant constructs. CHO cells were held at −90 mV and the membrane voltage was stepped for 3 s from −60 mV to +60 mV in 10 mV increments and then repolarized for 1.5 s to −60 mV. F, conductance-voltage relations of the WT Kv7.1 concatemer, L273F Kv7.1 monomer and L273F Kv7.1 concatemer. Data were fitted with a single Boltzmann function.

    Journal: Channels

    Article Title: Inactivation gating of Kv7.1 channels does not involve concerted cooperative subunit interactions

    doi: 10.1080/19336950.2018.1441649

    Figure Lengend Snippet: Characterization of the WT and L273F mutant Kv7.1 concatenated tetrameric channels. A, Top panel, scheme of the Kv7.1 concatenated tetrameric channel construct (Con), where subunits D1, D2, D3 and D4 are connected by flexible linkers. Each linker (8 glycines) harbors a unique restriction site. B, conductance-voltage relations of WT monomeric and WT concatenated tetrameric Kv7.1 constructs in the absence or presence of KCNE1. Data were fitted with a single Boltzmann function. C, Western blot showing lysates from HEK293 transfected with empty vector (Mock), KCNQ1 monomeric and concatenated tetrameric constructs. D, location of residue L273 in the S5 α helical segment of the Kv7.1 pore domain as mapped in the recent cryo-EM structure of the frog Kv7.1 (Sun, 2017). E, representative current traces of L273F Kv7.1 monomeric and concatenated tetrameric mutant constructs. CHO cells were held at −90 mV and the membrane voltage was stepped for 3 s from −60 mV to +60 mV in 10 mV increments and then repolarized for 1.5 s to −60 mV. F, conductance-voltage relations of the WT Kv7.1 concatemer, L273F Kv7.1 monomer and L273F Kv7.1 concatemer. Data were fitted with a single Boltzmann function.

    Article Snippet: Equal amounts of lysate proteins were resolved by 8% SDS-PAGE and blots were reacted using rabbit anti-Kv7.1 antibodies (Alomone Labs).

    Techniques: Mutagenesis, Construct, Western Blot, Transfection, Plasmid Preparation, Cryo-EM Sample Prep

    Recovery from inactivation of the different concatemer constructs. A, representative current traces of the recovery from inactivation of the different concatemer constructs. The recovery time from inactivation was determined from a −90 mV holding potential, by stepping the membrane voltage for 1.5 s to +50 mV to open the various Kv7.1 concatemer channels (first pulse), then repolarizing to −90 mV for various times to allow recovery from inactivation and stepping back to +50 mV for 0.5 s to reopen the channels (second pulse). B, The fractional recovery was calculated by the peak current of the second pulse divided by the peak of the first pulse. These ratios were plotted against the recovery times (n = 4). C, time constants of the recovery time from inactivation were calculated from an exponential fit (one way ANOVA, n = 4; * P

    Journal: Channels

    Article Title: Inactivation gating of Kv7.1 channels does not involve concerted cooperative subunit interactions

    doi: 10.1080/19336950.2018.1441649

    Figure Lengend Snippet: Recovery from inactivation of the different concatemer constructs. A, representative current traces of the recovery from inactivation of the different concatemer constructs. The recovery time from inactivation was determined from a −90 mV holding potential, by stepping the membrane voltage for 1.5 s to +50 mV to open the various Kv7.1 concatemer channels (first pulse), then repolarizing to −90 mV for various times to allow recovery from inactivation and stepping back to +50 mV for 0.5 s to reopen the channels (second pulse). B, The fractional recovery was calculated by the peak current of the second pulse divided by the peak of the first pulse. These ratios were plotted against the recovery times (n = 4). C, time constants of the recovery time from inactivation were calculated from an exponential fit (one way ANOVA, n = 4; * P

    Article Snippet: Equal amounts of lysate proteins were resolved by 8% SDS-PAGE and blots were reacted using rabbit anti-Kv7.1 antibodies (Alomone Labs).

    Techniques: Construct