anti cav1 3  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Name:
    Anti Cav1 3 CACNA1D extracellular Antibody
    Description:
    Anti Cav1 3 CACNA1D extracellular Antibody is directed against an extracellular epitope of the rat protein Anti CaV1 3 CACNA1D extracellular Antibody ACC 311 can be used in western blot and immunohistochemistry and live cell imaging applications It is specially suited to detect CaV1 3 channel in live cells It has been designed to recognize CaV1 3 from mouse rat and human samples
    Catalog Number:
    ACC-311
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Live Cell Imaging, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
    Buy from Supplier


    Structured Review

    Alomone Labs anti cav1 3
    Anti Cav1 3 CACNA1D extracellular Antibody
    Anti Cav1 3 CACNA1D extracellular Antibody is directed against an extracellular epitope of the rat protein Anti CaV1 3 CACNA1D extracellular Antibody ACC 311 can be used in western blot and immunohistochemistry and live cell imaging applications It is specially suited to detect CaV1 3 channel in live cells It has been designed to recognize CaV1 3 from mouse rat and human samples
    https://www.bioz.com/result/anti cav1 3/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 3 - by Bioz Stars, 2021-09
    92/100 stars

    Images

    1) Product Images from "Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells"

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    Journal: Journal of Neurochemistry

    doi: 10.1111/j.1471-4159.2010.07089.x

    Cav1 channel deletion compensated by the increased expression of other Cav channel types. Pharmacological dissection of Ca 2+ channels in mouse chromaffin cells from WT and Cav1.3 −/− cells. (a and b) Time course of the Ca 2+ charge density obtained after sequentially and cumulatively adding the different Ca 2+ channel blockers, in WT and Cav1.3 −/− cells, respectively: 3 μM nifedipine was used to block Cav1 channels, 1 μM ω‐CTX‐GVIA to block Cav2.2 channels, 3 μM ω‐CTX‐MVIIC to block Cav2.1 channels, and 200 μM Cd 2+ to block the residual Ca 2+ current. (c and d) Original traces of the Ca 2+ currents recorded at the stationary stage using each Ca 2+ channel blocker (corresponding to points a–f in panels 3a and b, where a : control, b : after 3 μM nifedipine perfusion, c : after 3 μM nifedipine and 1 μM ω‐CTX‐GVIA perfusion and d : after 3 μM nifedipine, ω‐CTX‐GVIA and 3 μM ω‐CTX‐MVIIC perfusion). (e) Ca 2+ charge density of the different Ca 2+ channel types for WT (black columns) and Cav1.3 −/− cells (white columns), respectively. (f) Total Ca 2+ charge obtained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). (g) Sizes of chromaffin cells obtained from WT (black column) and Cav1.3 −/− mice (white column). Experiments were performed on nine paired cultures of WT ( n = 18 cells) and Cav1.3 −/− cells ( n = 17 cells), using 1–2 mice of each strain. Bars represent means ± SEM. ** p
    Figure Legend Snippet: Cav1 channel deletion compensated by the increased expression of other Cav channel types. Pharmacological dissection of Ca 2+ channels in mouse chromaffin cells from WT and Cav1.3 −/− cells. (a and b) Time course of the Ca 2+ charge density obtained after sequentially and cumulatively adding the different Ca 2+ channel blockers, in WT and Cav1.3 −/− cells, respectively: 3 μM nifedipine was used to block Cav1 channels, 1 μM ω‐CTX‐GVIA to block Cav2.2 channels, 3 μM ω‐CTX‐MVIIC to block Cav2.1 channels, and 200 μM Cd 2+ to block the residual Ca 2+ current. (c and d) Original traces of the Ca 2+ currents recorded at the stationary stage using each Ca 2+ channel blocker (corresponding to points a–f in panels 3a and b, where a : control, b : after 3 μM nifedipine perfusion, c : after 3 μM nifedipine and 1 μM ω‐CTX‐GVIA perfusion and d : after 3 μM nifedipine, ω‐CTX‐GVIA and 3 μM ω‐CTX‐MVIIC perfusion). (e) Ca 2+ charge density of the different Ca 2+ channel types for WT (black columns) and Cav1.3 −/− cells (white columns), respectively. (f) Total Ca 2+ charge obtained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). (g) Sizes of chromaffin cells obtained from WT (black column) and Cav1.3 −/− mice (white column). Experiments were performed on nine paired cultures of WT ( n = 18 cells) and Cav1.3 −/− cells ( n = 17 cells), using 1–2 mice of each strain. Bars represent means ± SEM. ** p

    Techniques Used: Expressing, Dissection, Blocking Assay, Mouse Assay

    Contribution of Cav1 channel subtypes to pacemaking activity, and shaping of action potential waveform. (a–b) Recordings of the spontaneous firing of action potentials performed in the current clamp configuration in WT (a) or Cav1.3 −/− cells (b) under control conditions, and after perfusion with 300 nM nifedipine and 3 μM nifedipine. (c–d) Mean action potential obtained by averaging the action potentials recorded over 10 s under control conditions or after 300 nM and 3 μM nifedipine application to WT ( n = 6) (c) or Cav1.3 −/− cells ( n = 4) (d). Data were obtained in two paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain.
    Figure Legend Snippet: Contribution of Cav1 channel subtypes to pacemaking activity, and shaping of action potential waveform. (a–b) Recordings of the spontaneous firing of action potentials performed in the current clamp configuration in WT (a) or Cav1.3 −/− cells (b) under control conditions, and after perfusion with 300 nM nifedipine and 3 μM nifedipine. (c–d) Mean action potential obtained by averaging the action potentials recorded over 10 s under control conditions or after 300 nM and 3 μM nifedipine application to WT ( n = 6) (c) or Cav1.3 −/− cells ( n = 4) (d). Data were obtained in two paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain.

    Techniques Used: Activity Assay, Mouse Assay

    Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) Phase‐plane plot obtained by plotting dV/dt versus the voltage stimulus in WT and Cav1.3 −/− , respectively. Arrows indicate the points at which dV/dt increased from the initial baseline (threshold potential). Estimated threshold potentials were −27 mV and −22 mV for WT and Cav1.3 −/− cells, respectively; (c–d) action potential clamp experiments were performed by applying the mean voltage stimulus obtained under control conditions in Fig. 8(g and h) every 30 s. Starting from ‘Solution 3’ (see Material and Methods ), different blockers were sequentially added to that solution: 2 μM TTX (Solution 3 + TTX), 45 mM TEA (Solution 3 + TTX + TEA), 3 μM nifedipine (Solution 3 + TTX + TEA + Nife) and 200 μM CdCl 2 (Solution 3 + TTX + TEA + Nife + Cd). Perfusion with each solution was continued for at least 2 min so that the currents reached the steady‐state. Number of cell: 22 WT cells, 25 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain. (e–f) Ion currents were calculated from the recordings of panels (c) and (d), for WT and Cav1.3 −/− cells, respectively. To obtain the Na + current, the current obtained after perfusion with ‘Solution 3 + TTX’ was subtracted from the ‘Solution 3’ current. The K + current was calculated as the difference in the current yielded under ‘Solution 3 + TTX + TEA’ minus ‘Solution 3 + TTX’. The Cav1 current was obtained as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Nife’ and the total Ca 2+ current as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Cd’. The voltage stimulus, obtained from Fig. 8(g and h) , control conditions, was superimposed on the ion currents. Data were obtained in 4 paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.
    Figure Legend Snippet: Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) Phase‐plane plot obtained by plotting dV/dt versus the voltage stimulus in WT and Cav1.3 −/− , respectively. Arrows indicate the points at which dV/dt increased from the initial baseline (threshold potential). Estimated threshold potentials were −27 mV and −22 mV for WT and Cav1.3 −/− cells, respectively; (c–d) action potential clamp experiments were performed by applying the mean voltage stimulus obtained under control conditions in Fig. 8(g and h) every 30 s. Starting from ‘Solution 3’ (see Material and Methods ), different blockers were sequentially added to that solution: 2 μM TTX (Solution 3 + TTX), 45 mM TEA (Solution 3 + TTX + TEA), 3 μM nifedipine (Solution 3 + TTX + TEA + Nife) and 200 μM CdCl 2 (Solution 3 + TTX + TEA + Nife + Cd). Perfusion with each solution was continued for at least 2 min so that the currents reached the steady‐state. Number of cell: 22 WT cells, 25 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain. (e–f) Ion currents were calculated from the recordings of panels (c) and (d), for WT and Cav1.3 −/− cells, respectively. To obtain the Na + current, the current obtained after perfusion with ‘Solution 3 + TTX’ was subtracted from the ‘Solution 3’ current. The K + current was calculated as the difference in the current yielded under ‘Solution 3 + TTX + TEA’ minus ‘Solution 3 + TTX’. The Cav1 current was obtained as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Nife’ and the total Ca 2+ current as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Cd’. The voltage stimulus, obtained from Fig. 8(g and h) , control conditions, was superimposed on the ion currents. Data were obtained in 4 paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Techniques Used: Activity Assay, Mouse Assay

    Cav1 channel subtypes expressed in mouse chromaffin cells. Sensitivity of Cav1 channel subtypes to DHPs. Square‐step depolarizing pulses of 50 ms duration were applied every 30 s to the peak current voltage. (a, c and e) Ca 2+ charge density blockade obtained after perfusion with 300 nM (black columns) and 3 μM (white columns) nifedipine (Nife) in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. A large fraction of Cav1.3 −/− cells ( n = 21) did not respond to 300 nM nifedipine. (b, d and f) Original Ca 2+ current traces under control conditions or after perfusion with 300 nM or 3 μM nifedipine in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. Experiments were performed on seven paired cultures of WT and Cav1.3 −/− cells and five paired cultures of WT and Cav1.2DHP −/− cells, using two mice from each strain. Numbers of cells indicated in parentheses. Bars represent means ± SEM. *** p
    Figure Legend Snippet: Cav1 channel subtypes expressed in mouse chromaffin cells. Sensitivity of Cav1 channel subtypes to DHPs. Square‐step depolarizing pulses of 50 ms duration were applied every 30 s to the peak current voltage. (a, c and e) Ca 2+ charge density blockade obtained after perfusion with 300 nM (black columns) and 3 μM (white columns) nifedipine (Nife) in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. A large fraction of Cav1.3 −/− cells ( n = 21) did not respond to 300 nM nifedipine. (b, d and f) Original Ca 2+ current traces under control conditions or after perfusion with 300 nM or 3 μM nifedipine in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. Experiments were performed on seven paired cultures of WT and Cav1.3 −/− cells and five paired cultures of WT and Cav1.2DHP −/− cells, using two mice from each strain. Numbers of cells indicated in parentheses. Bars represent means ± SEM. *** p

    Techniques Used: Mouse Assay

    Voltage dependent activation of Ca 2+ channels. (a) I – V curves obtained under control conditions in WT and Cav1.3 −/− cells ( n = 8–12). 200 ms square‐step depolarizing pulses at increasing potentials (voltage increments of 10 mV), from −50 mV to 80 mV, were applied every 1 min. Data were obtained from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice from each strain and normalized as the percentage of current in control conditions at 10 mV, plotted as the mean ± SEM. * p
    Figure Legend Snippet: Voltage dependent activation of Ca 2+ channels. (a) I – V curves obtained under control conditions in WT and Cav1.3 −/− cells ( n = 8–12). 200 ms square‐step depolarizing pulses at increasing potentials (voltage increments of 10 mV), from −50 mV to 80 mV, were applied every 1 min. Data were obtained from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice from each strain and normalized as the percentage of current in control conditions at 10 mV, plotted as the mean ± SEM. * p

    Techniques Used: Activation Assay, Mouse Assay

    Cav1 channel deletion compensated by the increased expression of other Ca 2+ channel types. Contribution of Cav channels to the exocytosis of neurotransmitters in mouse chromaffin cells from WT and Cav1.3 −/− mice. (a and b) C m traces recorded simultaneously to the Ca 2+ currents of Fig. 2(c and d) in WT and Cav1.3 −/− cells, respectively. (c) Percentage of total secretion attributed to each Ca 2+ channel type in WT (black columns) and Cav1.3 −/− cells (white columns). (d) Total secretion attained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). Experiments were performed on seven paired cultures of WT ( n = 15 cells) and Cav1.3 −/− cells ( n = 11 cells), using 1–2 mice from each strain. Bars represent means ± SEM. * p
    Figure Legend Snippet: Cav1 channel deletion compensated by the increased expression of other Ca 2+ channel types. Contribution of Cav channels to the exocytosis of neurotransmitters in mouse chromaffin cells from WT and Cav1.3 −/− mice. (a and b) C m traces recorded simultaneously to the Ca 2+ currents of Fig. 2(c and d) in WT and Cav1.3 −/− cells, respectively. (c) Percentage of total secretion attributed to each Ca 2+ channel type in WT (black columns) and Cav1.3 −/− cells (white columns). (d) Total secretion attained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). Experiments were performed on seven paired cultures of WT ( n = 15 cells) and Cav1.3 −/− cells ( n = 11 cells), using 1–2 mice from each strain. Bars represent means ± SEM. * p

    Techniques Used: Expressing, Mouse Assay

    Contribution of Cav1 channel subtypes to the Ca 2+ charge and exocytosis of chromaffin vesicles. Ca 2+ charge density (a) and the corresponding exocytosis (b) versus the voltage in WT and Cav1.3 −/− cells. Data were obtained in the same experiments as in Fig. 3 ( n = 8–12, from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain) and normalized as the percentage of charge density in control conditions at 10 mV, plotted as the mean ± SEM. (c) Original traces of Ca 2+ current density and the corresponding exocytosis elicited at −20 mV, −10 mV and 0 mV in WT and Cav1.3 −/− cells. * p
    Figure Legend Snippet: Contribution of Cav1 channel subtypes to the Ca 2+ charge and exocytosis of chromaffin vesicles. Ca 2+ charge density (a) and the corresponding exocytosis (b) versus the voltage in WT and Cav1.3 −/− cells. Data were obtained in the same experiments as in Fig. 3 ( n = 8–12, from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain) and normalized as the percentage of charge density in control conditions at 10 mV, plotted as the mean ± SEM. (c) Original traces of Ca 2+ current density and the corresponding exocytosis elicited at −20 mV, −10 mV and 0 mV in WT and Cav1.3 −/− cells. * p

    Techniques Used: Mouse Assay

    Kinetics of the Cav1 channel subtypes. One‐second square‐step depolarizing pulses were applied at −10 mV every 5 min. (a) Inactivation kinetics. Left, Ca 2+ current remaining at the end of a 1‐s pulse expressed as a percentage of the peak current ( I 1000 / I peak ) in WT (black column) and Cav1.3 −/− cells (grey column); middle, percentage of cells whose inactivation kinetics could be well fitted to a single (τ inact single , black columns) or to a double (τ inact double , grey columns) exponential function in WT and Cav1.3 −/− ; right, the average τ inact single yielded by the single exponential fitting, and τ inact double , which exhibited two components, a fast component (τ inact fast ) and a slow component (τ inact slow ), were plotted for WT and Cav1.3 −/− cells (black and grey columns, respectively). (b) Original traces of the Cav1 channel currents recorded in WT and Cav1.3 −/− cells were averaged, superimposed and scaled to the peak WT Cav1 channel current. Number of cells indicated in parentheses. Data were obtained in three paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.
    Figure Legend Snippet: Kinetics of the Cav1 channel subtypes. One‐second square‐step depolarizing pulses were applied at −10 mV every 5 min. (a) Inactivation kinetics. Left, Ca 2+ current remaining at the end of a 1‐s pulse expressed as a percentage of the peak current ( I 1000 / I peak ) in WT (black column) and Cav1.3 −/− cells (grey column); middle, percentage of cells whose inactivation kinetics could be well fitted to a single (τ inact single , black columns) or to a double (τ inact double , grey columns) exponential function in WT and Cav1.3 −/− ; right, the average τ inact single yielded by the single exponential fitting, and τ inact double , which exhibited two components, a fast component (τ inact fast ) and a slow component (τ inact slow ), were plotted for WT and Cav1.3 −/− cells (black and grey columns, respectively). (b) Original traces of the Cav1 channel currents recorded in WT and Cav1.3 −/− cells were averaged, superimposed and scaled to the peak WT Cav1 channel current. Number of cells indicated in parentheses. Data were obtained in three paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Techniques Used: Mouse Assay

    Cav1 channel subtypes expressed in mouse chromaffin cells. Immunocytochemical characterization of Cav1 channel subtypes. (a–b) Confocal images of isolated mouse chromaffin cells from WT (a) or Cav1.3 −/− mice (b) labeled with antibodies against Cav1.1, Cav1.2, Cav1.3 and Cav1.4 channels (dilution 1 : 200) and the corresponding secondary antibody (dilution 1 : 200) Alexa Fluor excited at a wavelength of 594 nm (dilution 1 : 200). Experiments were performed on four paired cultures of WT and Cav1.3 −/− cells. Calibration bar: 75 microns.
    Figure Legend Snippet: Cav1 channel subtypes expressed in mouse chromaffin cells. Immunocytochemical characterization of Cav1 channel subtypes. (a–b) Confocal images of isolated mouse chromaffin cells from WT (a) or Cav1.3 −/− mice (b) labeled with antibodies against Cav1.1, Cav1.2, Cav1.3 and Cav1.4 channels (dilution 1 : 200) and the corresponding secondary antibody (dilution 1 : 200) Alexa Fluor excited at a wavelength of 594 nm (dilution 1 : 200). Experiments were performed on four paired cultures of WT and Cav1.3 −/− cells. Calibration bar: 75 microns.

    Techniques Used: Isolation, Mouse Assay, Labeling

    Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) In a different set of experiments performed under the current‐clamp configuration, the spontaneous oscillatory activity resistant to TTX, obtained in half the cells treated with this toxin, was reversibly abolished by 300 nM nifedipine in WT (a) or Cav1.3 −/− cells (b). In the other half of the cells, reversible blockade of spontaneous action potentials by 2 μM TTX in WT (c) or Cav1.3 −/− (d) cells was achieved. Data were obtained in two paired cultures of WT and Cav1.3 −/− cells using two mice of each strain.
    Figure Legend Snippet: Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) In a different set of experiments performed under the current‐clamp configuration, the spontaneous oscillatory activity resistant to TTX, obtained in half the cells treated with this toxin, was reversibly abolished by 300 nM nifedipine in WT (a) or Cav1.3 −/− cells (b). In the other half of the cells, reversible blockade of spontaneous action potentials by 2 μM TTX in WT (c) or Cav1.3 −/− (d) cells was achieved. Data were obtained in two paired cultures of WT and Cav1.3 −/− cells using two mice of each strain.

    Techniques Used: Activity Assay, Mouse Assay

    Coupling of Cav1 channel subtypes to BK channels. (a) Upper section: double‐pulse protocol used to recruit BK channels. This included a 400 ms test pulse ( V t ) to 140 mV or above that potential (trace 1), followed by a 10‐ms pre‐pulse applied at 0 mV before V t (trace 2). The Ca 2+ dependent K + currents activated using this protocol were BK channels. Lower section: original K + current traces recorded using the above protocol under control conditions and after perfusion with different K + channel blockers, added sequentially and cumulatively: first, 200 nM apamin, 100 nM charibdotoxin (ChTx), and finally 45 mM TEA. Pulses were applied every 2 min. Numbers of cells are indicated in parentheses. (b) The K + charge density was averaged and normalized for each condition with respect to the current in the absence of a pre‐pulse. (c–d) Effects of 3 μM nifedipine on BK channel currents in WT (c) and Cav1.3 −/− cells (d). Number of cells: 14 WT cells, 11 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.
    Figure Legend Snippet: Coupling of Cav1 channel subtypes to BK channels. (a) Upper section: double‐pulse protocol used to recruit BK channels. This included a 400 ms test pulse ( V t ) to 140 mV or above that potential (trace 1), followed by a 10‐ms pre‐pulse applied at 0 mV before V t (trace 2). The Ca 2+ dependent K + currents activated using this protocol were BK channels. Lower section: original K + current traces recorded using the above protocol under control conditions and after perfusion with different K + channel blockers, added sequentially and cumulatively: first, 200 nM apamin, 100 nM charibdotoxin (ChTx), and finally 45 mM TEA. Pulses were applied every 2 min. Numbers of cells are indicated in parentheses. (b) The K + charge density was averaged and normalized for each condition with respect to the current in the absence of a pre‐pulse. (c–d) Effects of 3 μM nifedipine on BK channel currents in WT (c) and Cav1.3 −/− cells (d). Number of cells: 14 WT cells, 11 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Techniques Used: Mouse Assay

    2) Product Images from "Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice"

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.102203

    Age-related expression of Cav1.3 in auditory pathway. ( A ) The immunofluorescence of CaV1.3 in the auditory cortex (green, magnification, ×400). ( B ) the quantitative analysis of CaV1.3 expression in the auditory cortex. ( C ) the western-blotting analysis of CaV1.3 expression in auditory cortex (top), the bottom panel is the quantitative analysis. ( D ) the mRNA expression of CaV1.3 in auditory cortex, inferior colliculus and cochlear nucleus. ( E ) CaV1.3 expression in auditory cortex was analyzed by flow cytometry, the right panel is the quantitative analysis. ( F ) Cav1.3 expression in neurons of auditory cortex, the neuron cells were gated as NeuN+, the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P
    Figure Legend Snippet: Age-related expression of Cav1.3 in auditory pathway. ( A ) The immunofluorescence of CaV1.3 in the auditory cortex (green, magnification, ×400). ( B ) the quantitative analysis of CaV1.3 expression in the auditory cortex. ( C ) the western-blotting analysis of CaV1.3 expression in auditory cortex (top), the bottom panel is the quantitative analysis. ( D ) the mRNA expression of CaV1.3 in auditory cortex, inferior colliculus and cochlear nucleus. ( E ) CaV1.3 expression in auditory cortex was analyzed by flow cytometry, the right panel is the quantitative analysis. ( F ) Cav1.3 expression in neurons of auditory cortex, the neuron cells were gated as NeuN+, the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Techniques Used: Expressing, Immunofluorescence, Western Blot, Flow Cytometry

    Age-related Cav1.3 expression in cochlea. ( A , B ) immunofluorescence of CaV1.3(green) and Myo7a (red) in the organ of Corti (left) and spiral ganglion (right) (magnification, ×400), nuclei was visualized by DAPI (blue). ( C ) the immunofluorescent staining for CaV1.3 (green) in the whole cochlear basilar membrane. ( D ) quantitative analysis of CaV1.3 expression in hair cells, spiral ganglion and cochlea basilar membrane.
    Figure Legend Snippet: Age-related Cav1.3 expression in cochlea. ( A , B ) immunofluorescence of CaV1.3(green) and Myo7a (red) in the organ of Corti (left) and spiral ganglion (right) (magnification, ×400), nuclei was visualized by DAPI (blue). ( C ) the immunofluorescent staining for CaV1.3 (green) in the whole cochlear basilar membrane. ( D ) quantitative analysis of CaV1.3 expression in hair cells, spiral ganglion and cochlea basilar membrane.

    Techniques Used: Expressing, Immunofluorescence, Staining

    Hair cells were vulnerable to ROS injury after Cav1.3 was knocked out. ( A ) the effect of CaV1.3 knock out in HEI-OC1 was analyzed by flow cytometry. ( B ) membrane potential (top) and non-linear capacitance (NLC) (bottom) studies in WT HEI-OC1 and CaV1.3 KO HEI-OC1 cells (n=5). ( C ) western-blotting analysis of CaV1.3 and p53 expression in control and senescence HEI-OC1 cells induced by D-galactose (D-Gal) or hydrogen peroxide (H 2 O 2 ), the bottom panel is the quantitative analysis. ( D ) β-Galactosidase staining (top) and C12FDG staining (bottom) of control and senescent HEI-OC1 cells induced by H 2 O 2 . ( E ) flow cytometry analysis of CaV1.3 in control and H 2 O 2 induced HEI-OC1 cells. ( F ) C12FDG staining (top) and β-Galactosidase staining (bottom) of NC (negative control) and KO (CaV1.3 knock out) HEI-OC1 cells after H 2 O 2 induction. ( G, H ) CFSE staining and red dot staining of NC and KO HEI-OC1 cells with or without H 2 O 2 induction. ( I ) LDH assay of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). ( J ) caspase-3/7-AAD staining of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). Error bars represent mean ± s.d.; *P
    Figure Legend Snippet: Hair cells were vulnerable to ROS injury after Cav1.3 was knocked out. ( A ) the effect of CaV1.3 knock out in HEI-OC1 was analyzed by flow cytometry. ( B ) membrane potential (top) and non-linear capacitance (NLC) (bottom) studies in WT HEI-OC1 and CaV1.3 KO HEI-OC1 cells (n=5). ( C ) western-blotting analysis of CaV1.3 and p53 expression in control and senescence HEI-OC1 cells induced by D-galactose (D-Gal) or hydrogen peroxide (H 2 O 2 ), the bottom panel is the quantitative analysis. ( D ) β-Galactosidase staining (top) and C12FDG staining (bottom) of control and senescent HEI-OC1 cells induced by H 2 O 2 . ( E ) flow cytometry analysis of CaV1.3 in control and H 2 O 2 induced HEI-OC1 cells. ( F ) C12FDG staining (top) and β-Galactosidase staining (bottom) of NC (negative control) and KO (CaV1.3 knock out) HEI-OC1 cells after H 2 O 2 induction. ( G, H ) CFSE staining and red dot staining of NC and KO HEI-OC1 cells with or without H 2 O 2 induction. ( I ) LDH assay of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). ( J ) caspase-3/7-AAD staining of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). Error bars represent mean ± s.d.; *P

    Techniques Used: Knock-Out, Flow Cytometry, Western Blot, Expressing, Staining, Negative Control, Lactate Dehydrogenase Assay

    Cav1.3 knock out decrease intra cellular calcium and subsequently result in reduction of complex I derived ROS inactivation. ( A ) intra-cellular ROS detection by flow cytometry (n=3). ( B – E ) the intra cellular calcium, intra cellular ROS, caspase-3/7-AAD staining and C12FDG staining of NC and KO HEI-OC1 cells with or without Ionmycin (n=3). ( F ) immunofluorescence of mitoSOX (red) and mitotrackor (green) in NC and KO HEI-OC1 cells, nuclei was visualized by DAPI (magnification, ×400, scal bar: 50μm), the right panels are the quantitative analysis of mitoSOX (top) and mitoTrackor (bottom). ( G ) the intra cellular ROS of KO HEI-OC1 cells with or without gradient Retenone (Ret) and Antimycin A (AMA)(n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P
    Figure Legend Snippet: Cav1.3 knock out decrease intra cellular calcium and subsequently result in reduction of complex I derived ROS inactivation. ( A ) intra-cellular ROS detection by flow cytometry (n=3). ( B – E ) the intra cellular calcium, intra cellular ROS, caspase-3/7-AAD staining and C12FDG staining of NC and KO HEI-OC1 cells with or without Ionmycin (n=3). ( F ) immunofluorescence of mitoSOX (red) and mitotrackor (green) in NC and KO HEI-OC1 cells, nuclei was visualized by DAPI (magnification, ×400, scal bar: 50μm), the right panels are the quantitative analysis of mitoSOX (top) and mitoTrackor (bottom). ( G ) the intra cellular ROS of KO HEI-OC1 cells with or without gradient Retenone (Ret) and Antimycin A (AMA)(n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Techniques Used: Knock-Out, Derivative Assay, Flow Cytometry, Staining, Immunofluorescence

    Cav1.3 knock down aggravated the loss of hair cells after senescence induction and resulted in hearing impairment. ( A ) immunofluorescence of CaV1.3 (green) and mCherry (red) in the organ of Corti (left) and spiral ganglion (right) of control and CaV1.3 knock down AAV group, nuclei was visualized by DAPI (magnification, ×400, scale bar: 50μm), the right panels are the quantitative analysis of CaV1.3 expression in organ of corti (top) and spiral ganglion (bottom). ( B ) auditory brainstem response (ABR) (top) and the whole cochlear basilar membrane after DAPI staining (bottom) of NC, CaV1.3 knock down, NC+D-Gal and CaV1.3 knock down+D-Gal group (n=6). ( C ) phalloidine staining for control and negative control AAV or Cav1.3 knock down AAV infected OC segment explants with H 2 O 2 treatment (n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P
    Figure Legend Snippet: Cav1.3 knock down aggravated the loss of hair cells after senescence induction and resulted in hearing impairment. ( A ) immunofluorescence of CaV1.3 (green) and mCherry (red) in the organ of Corti (left) and spiral ganglion (right) of control and CaV1.3 knock down AAV group, nuclei was visualized by DAPI (magnification, ×400, scale bar: 50μm), the right panels are the quantitative analysis of CaV1.3 expression in organ of corti (top) and spiral ganglion (bottom). ( B ) auditory brainstem response (ABR) (top) and the whole cochlear basilar membrane after DAPI staining (bottom) of NC, CaV1.3 knock down, NC+D-Gal and CaV1.3 knock down+D-Gal group (n=6). ( C ) phalloidine staining for control and negative control AAV or Cav1.3 knock down AAV infected OC segment explants with H 2 O 2 treatment (n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Techniques Used: Immunofluorescence, Expressing, Staining, Negative Control, Infection

    Related Articles

    Produced:

    Article Title: Distinct localization and modulation of Cav1.2 and Cav1.3 L-type Ca2+ channels in mouse sinoatrial node
    Article Snippet: .. In some experiments, commercially available rabbit polyclonal Cav 1.3 antibodies (Alomone Labs, Jerusalem, Israel) were used, which produced qualitatively similar results as the Cav1.3 antibodies we characterized previously ( ). ..

    Incubation:

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice
    Article Snippet: .. After overnight incubation with the primary antibody, rabbit anti-CaV1.3 calcium channel polyclonal antibody (1:50; Alomone labs, Israel) or mouse anti-Myo7a antibody (1:100; Santa Cruz, CA) at 4°C the sections were washed three times with PBS and incubated using Dylight 488 conjugated goat anti-rabbit IgG or Dylight 594 conjugated goat anti-mouse (1:500; Multi-Sciences, Hangzhou, China) for 1 h at room temperature. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Alomone Labs anti cav1 3
    Cav1 channel deletion compensated by the increased expression of other Cav channel types. Pharmacological dissection of Ca 2+ channels in mouse chromaffin cells from WT and <t>Cav1.3</t> −/− cells. (a and b) Time course of the Ca 2+ charge density obtained after sequentially and cumulatively adding the different Ca 2+ channel blockers, in WT and Cav1.3 −/− cells, respectively: 3 μM nifedipine was used to block Cav1 channels, 1 μM ω‐CTX‐GVIA to block Cav2.2 channels, 3 μM ω‐CTX‐MVIIC to block Cav2.1 channels, and 200 μM Cd 2+ to block the residual Ca 2+ current. (c and d) Original traces of the Ca 2+ currents recorded at the stationary stage using each Ca 2+ channel blocker (corresponding to points a–f in panels 3a and b, where a : control, b : after 3 μM nifedipine perfusion, c : after 3 μM nifedipine and 1 μM ω‐CTX‐GVIA perfusion and d : after 3 μM nifedipine, ω‐CTX‐GVIA and 3 μM ω‐CTX‐MVIIC perfusion). (e) Ca 2+ charge density of the different Ca 2+ channel types for WT (black columns) and Cav1.3 −/− cells (white columns), respectively. (f) Total Ca 2+ charge obtained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). (g) Sizes of chromaffin cells obtained from WT (black column) and Cav1.3 −/− mice (white column). Experiments were performed on nine paired cultures of WT ( n = 18 cells) and Cav1.3 −/− cells ( n = 17 cells), using 1–2 mice of each strain. Bars represent means ± SEM. ** p
    Anti Cav1 3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cav1 3/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 3 - by Bioz Stars, 2021-09
    92/100 stars
      Buy from Supplier

    Image Search Results


    Cav1 channel deletion compensated by the increased expression of other Cav channel types. Pharmacological dissection of Ca 2+ channels in mouse chromaffin cells from WT and Cav1.3 −/− cells. (a and b) Time course of the Ca 2+ charge density obtained after sequentially and cumulatively adding the different Ca 2+ channel blockers, in WT and Cav1.3 −/− cells, respectively: 3 μM nifedipine was used to block Cav1 channels, 1 μM ω‐CTX‐GVIA to block Cav2.2 channels, 3 μM ω‐CTX‐MVIIC to block Cav2.1 channels, and 200 μM Cd 2+ to block the residual Ca 2+ current. (c and d) Original traces of the Ca 2+ currents recorded at the stationary stage using each Ca 2+ channel blocker (corresponding to points a–f in panels 3a and b, where a : control, b : after 3 μM nifedipine perfusion, c : after 3 μM nifedipine and 1 μM ω‐CTX‐GVIA perfusion and d : after 3 μM nifedipine, ω‐CTX‐GVIA and 3 μM ω‐CTX‐MVIIC perfusion). (e) Ca 2+ charge density of the different Ca 2+ channel types for WT (black columns) and Cav1.3 −/− cells (white columns), respectively. (f) Total Ca 2+ charge obtained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). (g) Sizes of chromaffin cells obtained from WT (black column) and Cav1.3 −/− mice (white column). Experiments were performed on nine paired cultures of WT ( n = 18 cells) and Cav1.3 −/− cells ( n = 17 cells), using 1–2 mice of each strain. Bars represent means ± SEM. ** p

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Cav1 channel deletion compensated by the increased expression of other Cav channel types. Pharmacological dissection of Ca 2+ channels in mouse chromaffin cells from WT and Cav1.3 −/− cells. (a and b) Time course of the Ca 2+ charge density obtained after sequentially and cumulatively adding the different Ca 2+ channel blockers, in WT and Cav1.3 −/− cells, respectively: 3 μM nifedipine was used to block Cav1 channels, 1 μM ω‐CTX‐GVIA to block Cav2.2 channels, 3 μM ω‐CTX‐MVIIC to block Cav2.1 channels, and 200 μM Cd 2+ to block the residual Ca 2+ current. (c and d) Original traces of the Ca 2+ currents recorded at the stationary stage using each Ca 2+ channel blocker (corresponding to points a–f in panels 3a and b, where a : control, b : after 3 μM nifedipine perfusion, c : after 3 μM nifedipine and 1 μM ω‐CTX‐GVIA perfusion and d : after 3 μM nifedipine, ω‐CTX‐GVIA and 3 μM ω‐CTX‐MVIIC perfusion). (e) Ca 2+ charge density of the different Ca 2+ channel types for WT (black columns) and Cav1.3 −/− cells (white columns), respectively. (f) Total Ca 2+ charge obtained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). (g) Sizes of chromaffin cells obtained from WT (black column) and Cav1.3 −/− mice (white column). Experiments were performed on nine paired cultures of WT ( n = 18 cells) and Cav1.3 −/− cells ( n = 17 cells), using 1–2 mice of each strain. Bars represent means ± SEM. ** p

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Dissection, Blocking Assay, Mouse Assay

    Contribution of Cav1 channel subtypes to pacemaking activity, and shaping of action potential waveform. (a–b) Recordings of the spontaneous firing of action potentials performed in the current clamp configuration in WT (a) or Cav1.3 −/− cells (b) under control conditions, and after perfusion with 300 nM nifedipine and 3 μM nifedipine. (c–d) Mean action potential obtained by averaging the action potentials recorded over 10 s under control conditions or after 300 nM and 3 μM nifedipine application to WT ( n = 6) (c) or Cav1.3 −/− cells ( n = 4) (d). Data were obtained in two paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain.

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Contribution of Cav1 channel subtypes to pacemaking activity, and shaping of action potential waveform. (a–b) Recordings of the spontaneous firing of action potentials performed in the current clamp configuration in WT (a) or Cav1.3 −/− cells (b) under control conditions, and after perfusion with 300 nM nifedipine and 3 μM nifedipine. (c–d) Mean action potential obtained by averaging the action potentials recorded over 10 s under control conditions or after 300 nM and 3 μM nifedipine application to WT ( n = 6) (c) or Cav1.3 −/− cells ( n = 4) (d). Data were obtained in two paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain.

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Activity Assay, Mouse Assay

    Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) Phase‐plane plot obtained by plotting dV/dt versus the voltage stimulus in WT and Cav1.3 −/− , respectively. Arrows indicate the points at which dV/dt increased from the initial baseline (threshold potential). Estimated threshold potentials were −27 mV and −22 mV for WT and Cav1.3 −/− cells, respectively; (c–d) action potential clamp experiments were performed by applying the mean voltage stimulus obtained under control conditions in Fig. 8(g and h) every 30 s. Starting from ‘Solution 3’ (see Material and Methods ), different blockers were sequentially added to that solution: 2 μM TTX (Solution 3 + TTX), 45 mM TEA (Solution 3 + TTX + TEA), 3 μM nifedipine (Solution 3 + TTX + TEA + Nife) and 200 μM CdCl 2 (Solution 3 + TTX + TEA + Nife + Cd). Perfusion with each solution was continued for at least 2 min so that the currents reached the steady‐state. Number of cell: 22 WT cells, 25 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain. (e–f) Ion currents were calculated from the recordings of panels (c) and (d), for WT and Cav1.3 −/− cells, respectively. To obtain the Na + current, the current obtained after perfusion with ‘Solution 3 + TTX’ was subtracted from the ‘Solution 3’ current. The K + current was calculated as the difference in the current yielded under ‘Solution 3 + TTX + TEA’ minus ‘Solution 3 + TTX’. The Cav1 current was obtained as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Nife’ and the total Ca 2+ current as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Cd’. The voltage stimulus, obtained from Fig. 8(g and h) , control conditions, was superimposed on the ion currents. Data were obtained in 4 paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) Phase‐plane plot obtained by plotting dV/dt versus the voltage stimulus in WT and Cav1.3 −/− , respectively. Arrows indicate the points at which dV/dt increased from the initial baseline (threshold potential). Estimated threshold potentials were −27 mV and −22 mV for WT and Cav1.3 −/− cells, respectively; (c–d) action potential clamp experiments were performed by applying the mean voltage stimulus obtained under control conditions in Fig. 8(g and h) every 30 s. Starting from ‘Solution 3’ (see Material and Methods ), different blockers were sequentially added to that solution: 2 μM TTX (Solution 3 + TTX), 45 mM TEA (Solution 3 + TTX + TEA), 3 μM nifedipine (Solution 3 + TTX + TEA + Nife) and 200 μM CdCl 2 (Solution 3 + TTX + TEA + Nife + Cd). Perfusion with each solution was continued for at least 2 min so that the currents reached the steady‐state. Number of cell: 22 WT cells, 25 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain. (e–f) Ion currents were calculated from the recordings of panels (c) and (d), for WT and Cav1.3 −/− cells, respectively. To obtain the Na + current, the current obtained after perfusion with ‘Solution 3 + TTX’ was subtracted from the ‘Solution 3’ current. The K + current was calculated as the difference in the current yielded under ‘Solution 3 + TTX + TEA’ minus ‘Solution 3 + TTX’. The Cav1 current was obtained as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Nife’ and the total Ca 2+ current as the difference between ‘Solution 3 + TTX + TEA’ and ‘Solution 3 + TTX + TEA + Cd’. The voltage stimulus, obtained from Fig. 8(g and h) , control conditions, was superimposed on the ion currents. Data were obtained in 4 paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Activity Assay, Mouse Assay

    Cav1 channel subtypes expressed in mouse chromaffin cells. Sensitivity of Cav1 channel subtypes to DHPs. Square‐step depolarizing pulses of 50 ms duration were applied every 30 s to the peak current voltage. (a, c and e) Ca 2+ charge density blockade obtained after perfusion with 300 nM (black columns) and 3 μM (white columns) nifedipine (Nife) in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. A large fraction of Cav1.3 −/− cells ( n = 21) did not respond to 300 nM nifedipine. (b, d and f) Original Ca 2+ current traces under control conditions or after perfusion with 300 nM or 3 μM nifedipine in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. Experiments were performed on seven paired cultures of WT and Cav1.3 −/− cells and five paired cultures of WT and Cav1.2DHP −/− cells, using two mice from each strain. Numbers of cells indicated in parentheses. Bars represent means ± SEM. *** p

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Cav1 channel subtypes expressed in mouse chromaffin cells. Sensitivity of Cav1 channel subtypes to DHPs. Square‐step depolarizing pulses of 50 ms duration were applied every 30 s to the peak current voltage. (a, c and e) Ca 2+ charge density blockade obtained after perfusion with 300 nM (black columns) and 3 μM (white columns) nifedipine (Nife) in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. A large fraction of Cav1.3 −/− cells ( n = 21) did not respond to 300 nM nifedipine. (b, d and f) Original Ca 2+ current traces under control conditions or after perfusion with 300 nM or 3 μM nifedipine in WT, Cav1.3 −/− and Cav1.2DHP −/− cells, respectively. Experiments were performed on seven paired cultures of WT and Cav1.3 −/− cells and five paired cultures of WT and Cav1.2DHP −/− cells, using two mice from each strain. Numbers of cells indicated in parentheses. Bars represent means ± SEM. *** p

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Mouse Assay

    Voltage dependent activation of Ca 2+ channels. (a) I – V curves obtained under control conditions in WT and Cav1.3 −/− cells ( n = 8–12). 200 ms square‐step depolarizing pulses at increasing potentials (voltage increments of 10 mV), from −50 mV to 80 mV, were applied every 1 min. Data were obtained from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice from each strain and normalized as the percentage of current in control conditions at 10 mV, plotted as the mean ± SEM. * p

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Voltage dependent activation of Ca 2+ channels. (a) I – V curves obtained under control conditions in WT and Cav1.3 −/− cells ( n = 8–12). 200 ms square‐step depolarizing pulses at increasing potentials (voltage increments of 10 mV), from −50 mV to 80 mV, were applied every 1 min. Data were obtained from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice from each strain and normalized as the percentage of current in control conditions at 10 mV, plotted as the mean ± SEM. * p

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Activation Assay, Mouse Assay

    Cav1 channel deletion compensated by the increased expression of other Ca 2+ channel types. Contribution of Cav channels to the exocytosis of neurotransmitters in mouse chromaffin cells from WT and Cav1.3 −/− mice. (a and b) C m traces recorded simultaneously to the Ca 2+ currents of Fig. 2(c and d) in WT and Cav1.3 −/− cells, respectively. (c) Percentage of total secretion attributed to each Ca 2+ channel type in WT (black columns) and Cav1.3 −/− cells (white columns). (d) Total secretion attained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). Experiments were performed on seven paired cultures of WT ( n = 15 cells) and Cav1.3 −/− cells ( n = 11 cells), using 1–2 mice from each strain. Bars represent means ± SEM. * p

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Cav1 channel deletion compensated by the increased expression of other Ca 2+ channel types. Contribution of Cav channels to the exocytosis of neurotransmitters in mouse chromaffin cells from WT and Cav1.3 −/− mice. (a and b) C m traces recorded simultaneously to the Ca 2+ currents of Fig. 2(c and d) in WT and Cav1.3 −/− cells, respectively. (c) Percentage of total secretion attributed to each Ca 2+ channel type in WT (black columns) and Cav1.3 −/− cells (white columns). (d) Total secretion attained under control conditions for WT (black column) and Cav1.3 −/− cells (white column). Experiments were performed on seven paired cultures of WT ( n = 15 cells) and Cav1.3 −/− cells ( n = 11 cells), using 1–2 mice from each strain. Bars represent means ± SEM. * p

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Mouse Assay

    Contribution of Cav1 channel subtypes to the Ca 2+ charge and exocytosis of chromaffin vesicles. Ca 2+ charge density (a) and the corresponding exocytosis (b) versus the voltage in WT and Cav1.3 −/− cells. Data were obtained in the same experiments as in Fig. 3 ( n = 8–12, from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain) and normalized as the percentage of charge density in control conditions at 10 mV, plotted as the mean ± SEM. (c) Original traces of Ca 2+ current density and the corresponding exocytosis elicited at −20 mV, −10 mV and 0 mV in WT and Cav1.3 −/− cells. * p

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Contribution of Cav1 channel subtypes to the Ca 2+ charge and exocytosis of chromaffin vesicles. Ca 2+ charge density (a) and the corresponding exocytosis (b) versus the voltage in WT and Cav1.3 −/− cells. Data were obtained in the same experiments as in Fig. 3 ( n = 8–12, from four paired cultures of WT and Cav1.3 −/− cells, using 1–2 mice of each strain) and normalized as the percentage of charge density in control conditions at 10 mV, plotted as the mean ± SEM. (c) Original traces of Ca 2+ current density and the corresponding exocytosis elicited at −20 mV, −10 mV and 0 mV in WT and Cav1.3 −/− cells. * p

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Mouse Assay

    Kinetics of the Cav1 channel subtypes. One‐second square‐step depolarizing pulses were applied at −10 mV every 5 min. (a) Inactivation kinetics. Left, Ca 2+ current remaining at the end of a 1‐s pulse expressed as a percentage of the peak current ( I 1000 / I peak ) in WT (black column) and Cav1.3 −/− cells (grey column); middle, percentage of cells whose inactivation kinetics could be well fitted to a single (τ inact single , black columns) or to a double (τ inact double , grey columns) exponential function in WT and Cav1.3 −/− ; right, the average τ inact single yielded by the single exponential fitting, and τ inact double , which exhibited two components, a fast component (τ inact fast ) and a slow component (τ inact slow ), were plotted for WT and Cav1.3 −/− cells (black and grey columns, respectively). (b) Original traces of the Cav1 channel currents recorded in WT and Cav1.3 −/− cells were averaged, superimposed and scaled to the peak WT Cav1 channel current. Number of cells indicated in parentheses. Data were obtained in three paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Kinetics of the Cav1 channel subtypes. One‐second square‐step depolarizing pulses were applied at −10 mV every 5 min. (a) Inactivation kinetics. Left, Ca 2+ current remaining at the end of a 1‐s pulse expressed as a percentage of the peak current ( I 1000 / I peak ) in WT (black column) and Cav1.3 −/− cells (grey column); middle, percentage of cells whose inactivation kinetics could be well fitted to a single (τ inact single , black columns) or to a double (τ inact double , grey columns) exponential function in WT and Cav1.3 −/− ; right, the average τ inact single yielded by the single exponential fitting, and τ inact double , which exhibited two components, a fast component (τ inact fast ) and a slow component (τ inact slow ), were plotted for WT and Cav1.3 −/− cells (black and grey columns, respectively). (b) Original traces of the Cav1 channel currents recorded in WT and Cav1.3 −/− cells were averaged, superimposed and scaled to the peak WT Cav1 channel current. Number of cells indicated in parentheses. Data were obtained in three paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Mouse Assay

    Cav1 channel subtypes expressed in mouse chromaffin cells. Immunocytochemical characterization of Cav1 channel subtypes. (a–b) Confocal images of isolated mouse chromaffin cells from WT (a) or Cav1.3 −/− mice (b) labeled with antibodies against Cav1.1, Cav1.2, Cav1.3 and Cav1.4 channels (dilution 1 : 200) and the corresponding secondary antibody (dilution 1 : 200) Alexa Fluor excited at a wavelength of 594 nm (dilution 1 : 200). Experiments were performed on four paired cultures of WT and Cav1.3 −/− cells. Calibration bar: 75 microns.

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Cav1 channel subtypes expressed in mouse chromaffin cells. Immunocytochemical characterization of Cav1 channel subtypes. (a–b) Confocal images of isolated mouse chromaffin cells from WT (a) or Cav1.3 −/− mice (b) labeled with antibodies against Cav1.1, Cav1.2, Cav1.3 and Cav1.4 channels (dilution 1 : 200) and the corresponding secondary antibody (dilution 1 : 200) Alexa Fluor excited at a wavelength of 594 nm (dilution 1 : 200). Experiments were performed on four paired cultures of WT and Cav1.3 −/− cells. Calibration bar: 75 microns.

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Isolation, Mouse Assay, Labeling

    Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) In a different set of experiments performed under the current‐clamp configuration, the spontaneous oscillatory activity resistant to TTX, obtained in half the cells treated with this toxin, was reversibly abolished by 300 nM nifedipine in WT (a) or Cav1.3 −/− cells (b). In the other half of the cells, reversible blockade of spontaneous action potentials by 2 μM TTX in WT (c) or Cav1.3 −/− (d) cells was achieved. Data were obtained in two paired cultures of WT and Cav1.3 −/− cells using two mice of each strain.

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Contribution of Cav1 channel subtypes to pacemaking activity. (a–b) In a different set of experiments performed under the current‐clamp configuration, the spontaneous oscillatory activity resistant to TTX, obtained in half the cells treated with this toxin, was reversibly abolished by 300 nM nifedipine in WT (a) or Cav1.3 −/− cells (b). In the other half of the cells, reversible blockade of spontaneous action potentials by 2 μM TTX in WT (c) or Cav1.3 −/− (d) cells was achieved. Data were obtained in two paired cultures of WT and Cav1.3 −/− cells using two mice of each strain.

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Activity Assay, Mouse Assay

    Coupling of Cav1 channel subtypes to BK channels. (a) Upper section: double‐pulse protocol used to recruit BK channels. This included a 400 ms test pulse ( V t ) to 140 mV or above that potential (trace 1), followed by a 10‐ms pre‐pulse applied at 0 mV before V t (trace 2). The Ca 2+ dependent K + currents activated using this protocol were BK channels. Lower section: original K + current traces recorded using the above protocol under control conditions and after perfusion with different K + channel blockers, added sequentially and cumulatively: first, 200 nM apamin, 100 nM charibdotoxin (ChTx), and finally 45 mM TEA. Pulses were applied every 2 min. Numbers of cells are indicated in parentheses. (b) The K + charge density was averaged and normalized for each condition with respect to the current in the absence of a pre‐pulse. (c–d) Effects of 3 μM nifedipine on BK channel currents in WT (c) and Cav1.3 −/− cells (d). Number of cells: 14 WT cells, 11 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Journal: Journal of Neurochemistry

    Article Title: Different roles attributed to Cav1 channel subtypes in spontaneous action potential firing and fine tuning of exocytosis in mouse chromaffin cells

    doi: 10.1111/j.1471-4159.2010.07089.x

    Figure Lengend Snippet: Coupling of Cav1 channel subtypes to BK channels. (a) Upper section: double‐pulse protocol used to recruit BK channels. This included a 400 ms test pulse ( V t ) to 140 mV or above that potential (trace 1), followed by a 10‐ms pre‐pulse applied at 0 mV before V t (trace 2). The Ca 2+ dependent K + currents activated using this protocol were BK channels. Lower section: original K + current traces recorded using the above protocol under control conditions and after perfusion with different K + channel blockers, added sequentially and cumulatively: first, 200 nM apamin, 100 nM charibdotoxin (ChTx), and finally 45 mM TEA. Pulses were applied every 2 min. Numbers of cells are indicated in parentheses. (b) The K + charge density was averaged and normalized for each condition with respect to the current in the absence of a pre‐pulse. (c–d) Effects of 3 μM nifedipine on BK channel currents in WT (c) and Cav1.3 −/− cells (d). Number of cells: 14 WT cells, 11 Cav1.3 −/− cells. Data were obtained in four paired cultures of WT and Cav1.3 −/− cells, using two mice of each strain.

    Article Snippet: The primary antibodies (dilution 1 : 200) were goat polyclonal anti‐Cav1.1 and anti‐Cav1.4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit polyclonal anti‐Cav1.2 and anti‐Cav1.3 (Alomone Labs, Jerusalem, Israel).

    Techniques: Mouse Assay

    Age-related expression of Cav1.3 in auditory pathway. ( A ) The immunofluorescence of CaV1.3 in the auditory cortex (green, magnification, ×400). ( B ) the quantitative analysis of CaV1.3 expression in the auditory cortex. ( C ) the western-blotting analysis of CaV1.3 expression in auditory cortex (top), the bottom panel is the quantitative analysis. ( D ) the mRNA expression of CaV1.3 in auditory cortex, inferior colliculus and cochlear nucleus. ( E ) CaV1.3 expression in auditory cortex was analyzed by flow cytometry, the right panel is the quantitative analysis. ( F ) Cav1.3 expression in neurons of auditory cortex, the neuron cells were gated as NeuN+, the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Journal: Aging (Albany NY)

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice

    doi: 10.18632/aging.102203

    Figure Lengend Snippet: Age-related expression of Cav1.3 in auditory pathway. ( A ) The immunofluorescence of CaV1.3 in the auditory cortex (green, magnification, ×400). ( B ) the quantitative analysis of CaV1.3 expression in the auditory cortex. ( C ) the western-blotting analysis of CaV1.3 expression in auditory cortex (top), the bottom panel is the quantitative analysis. ( D ) the mRNA expression of CaV1.3 in auditory cortex, inferior colliculus and cochlear nucleus. ( E ) CaV1.3 expression in auditory cortex was analyzed by flow cytometry, the right panel is the quantitative analysis. ( F ) Cav1.3 expression in neurons of auditory cortex, the neuron cells were gated as NeuN+, the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Article Snippet: After overnight incubation with the primary antibody, rabbit anti-CaV1.3 calcium channel polyclonal antibody (1:50; Alomone labs, Israel) or mouse anti-Myo7a antibody (1:100; Santa Cruz, CA) at 4°C the sections were washed three times with PBS and incubated using Dylight 488 conjugated goat anti-rabbit IgG or Dylight 594 conjugated goat anti-mouse (1:500; Multi-Sciences, Hangzhou, China) for 1 h at room temperature.

    Techniques: Expressing, Immunofluorescence, Western Blot, Flow Cytometry

    Age-related Cav1.3 expression in cochlea. ( A , B ) immunofluorescence of CaV1.3(green) and Myo7a (red) in the organ of Corti (left) and spiral ganglion (right) (magnification, ×400), nuclei was visualized by DAPI (blue). ( C ) the immunofluorescent staining for CaV1.3 (green) in the whole cochlear basilar membrane. ( D ) quantitative analysis of CaV1.3 expression in hair cells, spiral ganglion and cochlea basilar membrane.

    Journal: Aging (Albany NY)

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice

    doi: 10.18632/aging.102203

    Figure Lengend Snippet: Age-related Cav1.3 expression in cochlea. ( A , B ) immunofluorescence of CaV1.3(green) and Myo7a (red) in the organ of Corti (left) and spiral ganglion (right) (magnification, ×400), nuclei was visualized by DAPI (blue). ( C ) the immunofluorescent staining for CaV1.3 (green) in the whole cochlear basilar membrane. ( D ) quantitative analysis of CaV1.3 expression in hair cells, spiral ganglion and cochlea basilar membrane.

    Article Snippet: After overnight incubation with the primary antibody, rabbit anti-CaV1.3 calcium channel polyclonal antibody (1:50; Alomone labs, Israel) or mouse anti-Myo7a antibody (1:100; Santa Cruz, CA) at 4°C the sections were washed three times with PBS and incubated using Dylight 488 conjugated goat anti-rabbit IgG or Dylight 594 conjugated goat anti-mouse (1:500; Multi-Sciences, Hangzhou, China) for 1 h at room temperature.

    Techniques: Expressing, Immunofluorescence, Staining

    Hair cells were vulnerable to ROS injury after Cav1.3 was knocked out. ( A ) the effect of CaV1.3 knock out in HEI-OC1 was analyzed by flow cytometry. ( B ) membrane potential (top) and non-linear capacitance (NLC) (bottom) studies in WT HEI-OC1 and CaV1.3 KO HEI-OC1 cells (n=5). ( C ) western-blotting analysis of CaV1.3 and p53 expression in control and senescence HEI-OC1 cells induced by D-galactose (D-Gal) or hydrogen peroxide (H 2 O 2 ), the bottom panel is the quantitative analysis. ( D ) β-Galactosidase staining (top) and C12FDG staining (bottom) of control and senescent HEI-OC1 cells induced by H 2 O 2 . ( E ) flow cytometry analysis of CaV1.3 in control and H 2 O 2 induced HEI-OC1 cells. ( F ) C12FDG staining (top) and β-Galactosidase staining (bottom) of NC (negative control) and KO (CaV1.3 knock out) HEI-OC1 cells after H 2 O 2 induction. ( G, H ) CFSE staining and red dot staining of NC and KO HEI-OC1 cells with or without H 2 O 2 induction. ( I ) LDH assay of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). ( J ) caspase-3/7-AAD staining of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). Error bars represent mean ± s.d.; *P

    Journal: Aging (Albany NY)

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice

    doi: 10.18632/aging.102203

    Figure Lengend Snippet: Hair cells were vulnerable to ROS injury after Cav1.3 was knocked out. ( A ) the effect of CaV1.3 knock out in HEI-OC1 was analyzed by flow cytometry. ( B ) membrane potential (top) and non-linear capacitance (NLC) (bottom) studies in WT HEI-OC1 and CaV1.3 KO HEI-OC1 cells (n=5). ( C ) western-blotting analysis of CaV1.3 and p53 expression in control and senescence HEI-OC1 cells induced by D-galactose (D-Gal) or hydrogen peroxide (H 2 O 2 ), the bottom panel is the quantitative analysis. ( D ) β-Galactosidase staining (top) and C12FDG staining (bottom) of control and senescent HEI-OC1 cells induced by H 2 O 2 . ( E ) flow cytometry analysis of CaV1.3 in control and H 2 O 2 induced HEI-OC1 cells. ( F ) C12FDG staining (top) and β-Galactosidase staining (bottom) of NC (negative control) and KO (CaV1.3 knock out) HEI-OC1 cells after H 2 O 2 induction. ( G, H ) CFSE staining and red dot staining of NC and KO HEI-OC1 cells with or without H 2 O 2 induction. ( I ) LDH assay of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). ( J ) caspase-3/7-AAD staining of NC and KO HEI-OC1 cells after H 2 O 2 induction (n=3). Error bars represent mean ± s.d.; *P

    Article Snippet: After overnight incubation with the primary antibody, rabbit anti-CaV1.3 calcium channel polyclonal antibody (1:50; Alomone labs, Israel) or mouse anti-Myo7a antibody (1:100; Santa Cruz, CA) at 4°C the sections were washed three times with PBS and incubated using Dylight 488 conjugated goat anti-rabbit IgG or Dylight 594 conjugated goat anti-mouse (1:500; Multi-Sciences, Hangzhou, China) for 1 h at room temperature.

    Techniques: Knock-Out, Flow Cytometry, Western Blot, Expressing, Staining, Negative Control, Lactate Dehydrogenase Assay

    Cav1.3 knock out decrease intra cellular calcium and subsequently result in reduction of complex I derived ROS inactivation. ( A ) intra-cellular ROS detection by flow cytometry (n=3). ( B – E ) the intra cellular calcium, intra cellular ROS, caspase-3/7-AAD staining and C12FDG staining of NC and KO HEI-OC1 cells with or without Ionmycin (n=3). ( F ) immunofluorescence of mitoSOX (red) and mitotrackor (green) in NC and KO HEI-OC1 cells, nuclei was visualized by DAPI (magnification, ×400, scal bar: 50μm), the right panels are the quantitative analysis of mitoSOX (top) and mitoTrackor (bottom). ( G ) the intra cellular ROS of KO HEI-OC1 cells with or without gradient Retenone (Ret) and Antimycin A (AMA)(n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Journal: Aging (Albany NY)

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice

    doi: 10.18632/aging.102203

    Figure Lengend Snippet: Cav1.3 knock out decrease intra cellular calcium and subsequently result in reduction of complex I derived ROS inactivation. ( A ) intra-cellular ROS detection by flow cytometry (n=3). ( B – E ) the intra cellular calcium, intra cellular ROS, caspase-3/7-AAD staining and C12FDG staining of NC and KO HEI-OC1 cells with or without Ionmycin (n=3). ( F ) immunofluorescence of mitoSOX (red) and mitotrackor (green) in NC and KO HEI-OC1 cells, nuclei was visualized by DAPI (magnification, ×400, scal bar: 50μm), the right panels are the quantitative analysis of mitoSOX (top) and mitoTrackor (bottom). ( G ) the intra cellular ROS of KO HEI-OC1 cells with or without gradient Retenone (Ret) and Antimycin A (AMA)(n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Article Snippet: After overnight incubation with the primary antibody, rabbit anti-CaV1.3 calcium channel polyclonal antibody (1:50; Alomone labs, Israel) or mouse anti-Myo7a antibody (1:100; Santa Cruz, CA) at 4°C the sections were washed three times with PBS and incubated using Dylight 488 conjugated goat anti-rabbit IgG or Dylight 594 conjugated goat anti-mouse (1:500; Multi-Sciences, Hangzhou, China) for 1 h at room temperature.

    Techniques: Knock-Out, Derivative Assay, Flow Cytometry, Staining, Immunofluorescence

    Cav1.3 knock down aggravated the loss of hair cells after senescence induction and resulted in hearing impairment. ( A ) immunofluorescence of CaV1.3 (green) and mCherry (red) in the organ of Corti (left) and spiral ganglion (right) of control and CaV1.3 knock down AAV group, nuclei was visualized by DAPI (magnification, ×400, scale bar: 50μm), the right panels are the quantitative analysis of CaV1.3 expression in organ of corti (top) and spiral ganglion (bottom). ( B ) auditory brainstem response (ABR) (top) and the whole cochlear basilar membrane after DAPI staining (bottom) of NC, CaV1.3 knock down, NC+D-Gal and CaV1.3 knock down+D-Gal group (n=6). ( C ) phalloidine staining for control and negative control AAV or Cav1.3 knock down AAV infected OC segment explants with H 2 O 2 treatment (n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Journal: Aging (Albany NY)

    Article Title: Down-regulation of Cav1.3 in auditory pathway promotes age-related hearing loss by enhancing calcium-mediated oxidative stress in male mice

    doi: 10.18632/aging.102203

    Figure Lengend Snippet: Cav1.3 knock down aggravated the loss of hair cells after senescence induction and resulted in hearing impairment. ( A ) immunofluorescence of CaV1.3 (green) and mCherry (red) in the organ of Corti (left) and spiral ganglion (right) of control and CaV1.3 knock down AAV group, nuclei was visualized by DAPI (magnification, ×400, scale bar: 50μm), the right panels are the quantitative analysis of CaV1.3 expression in organ of corti (top) and spiral ganglion (bottom). ( B ) auditory brainstem response (ABR) (top) and the whole cochlear basilar membrane after DAPI staining (bottom) of NC, CaV1.3 knock down, NC+D-Gal and CaV1.3 knock down+D-Gal group (n=6). ( C ) phalloidine staining for control and negative control AAV or Cav1.3 knock down AAV infected OC segment explants with H 2 O 2 treatment (n=3), the right panel is the quantitative analysis. Error bars represent mean ± s.d.; *P

    Article Snippet: After overnight incubation with the primary antibody, rabbit anti-CaV1.3 calcium channel polyclonal antibody (1:50; Alomone labs, Israel) or mouse anti-Myo7a antibody (1:100; Santa Cruz, CA) at 4°C the sections were washed three times with PBS and incubated using Dylight 488 conjugated goat anti-rabbit IgG or Dylight 594 conjugated goat anti-mouse (1:500; Multi-Sciences, Hangzhou, China) for 1 h at room temperature.

    Techniques: Immunofluorescence, Expressing, Staining, Negative Control, Infection