cay1 2 cacna1c antibody  (Alomone Labs)

Journal: Cells

Article Title: A Seven Amino Acid Long Region in the Cardiac Ca v 1.2, Encompassing Ser1487, Is Essential for Mice Embryonic Development and “Fight or Flight” Responses

doi: 10.3390/cells15181655

Figure Lengend Snippet: Full list of antibodies used in Western Blot experiments.

Article Snippet: CaV1.2 , rabbit , polyclonal , IgG , Alomone , ACC-003 , 1:200.

Techniques: Western Blot

Journal: bioRxiv

Article Title: Effect of ORL-1 on Cav1.2 calcium channels

doi: 10.64898/2026.07.03.736403

Figure Lengend Snippet: ORL-1 interacts with Cavα1c and Cavα2δ-1 of the Cav1.2 channel complex. a ) Co-immunoprecipitation from tsA-201 cell lysate, with immunoprecipitation for ORL-1-HIs and immunoblotting for Cavα1c (n = 5). Cavα1c co-immunoprecipitates with ORL-1 regardless of auxiliary subunit expression. b ) Immunoprecipitation for Cavα1c, with immunoblotting for ORL-1, demonstrating ORL-1 co-immunoprecipitates with Cavα1c in the presence and absence of auxiliary subunits (n = 3). c ) Immunoprecipitation for ORL-1-His, with immunoblotting for Cavα2δ-1, demonstrating that Cavα2δ-1 co-immunoprecipitates with ORL-1 when co-expressed both in the presence and absence of Cavα1c (n = 3).

Article Snippet: The volume equivalent to 1-2mg of protein was then incubated with anti-Xpress monoclonal antibody (Invitrogen) or anti-Cav1.2 (CACNA1C) antibody (Alomone CAT# ACC-003) at a ratio of 1-2 μg antibody per 1mg of protein.

Techniques: Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot, Expressing

Journal: bioRxiv

Article Title: Effect of ORL-1 on Cav1.2 calcium channels

doi: 10.64898/2026.07.03.736403

Figure Lengend Snippet: Co-expression of ORL-1 and Cav1.2 in tsA-201 cells decreases Cav1.2 peak current density without affecting biophysical properties. a) Representative whole cell traces for Cav1.2 and Cav1.2+ORL-1. Traces were evoked by depolarizing steps from -90 mV to +50 mV in 5 mV increments. b) Current-voltage relationships for Cav1.2 co-expressed with or without ORL-1 (n=21, n=14). All currents were normalized by cell capacitance. c) Half-activation voltage of Cav1.2 in the presence and the absence of ORL-1. d) Peak current density for Cav1.2 co-expressed with or without ORL-1. ORL-1 significantly reduced Cav1.2 peak current density (p = 0.0170, n=26, n=30). Peak current densities were taken from each individual trace, a majority of which occurred between -10 mV and 10 mV. Data are expressed as mean ± SEM.

Article Snippet: The volume equivalent to 1-2mg of protein was then incubated with anti-Xpress monoclonal antibody (Invitrogen) or anti-Cav1.2 (CACNA1C) antibody (Alomone CAT# ACC-003) at a ratio of 1-2 μg antibody per 1mg of protein.

Techniques: Expressing, Activation Assay, Patch Clamp

Journal: bioRxiv

Article Title: Effect of ORL-1 on Cav1.2 calcium channels

doi: 10.64898/2026.07.03.736403

Figure Lengend Snippet: Co-expression of ORL-1 and Cav1.2 does not alter Cav1.2 voltage dependence of inactivation or CDI. a) Steady-state inactivation curves for Cav1.2 co-expressed with or without ORL-1 in tsA-201 cells. ORL-1 did not significantly alter the slope or half-inactivation voltage. b) 2+ Calcium-dependent inactivation of Cav1.2 expressed with or without ORL-1 in 20 mM Ca external solutions, shown in the form of time constants of inactivation. Voltages around the average peak-current voltage (15 mV in 20 mM Ca 2+ ) were fitted with a single exponential. At all tested voltages, there was no difference between Cav1.2 alone or co-expressed with ORL-1. Data are expressed as mean ± SEM.

Article Snippet: The volume equivalent to 1-2mg of protein was then incubated with anti-Xpress monoclonal antibody (Invitrogen) or anti-Cav1.2 (CACNA1C) antibody (Alomone CAT# ACC-003) at a ratio of 1-2 μg antibody per 1mg of protein.

Techniques: Expressing, Inactivation

Journal: bioRxiv

Article Title: Effect of ORL-1 on Cav1.2 calcium channels

doi: 10.64898/2026.07.03.736403

Figure Lengend Snippet: Effect of nociceptin on Cav1.2 current amplitude. ORL-1 and Cav1.2 were co-expressed in tsA-201 cells and treated with acute continuous perfusion of external solution (VEH), followed by 1 µM nociceptin dissolved in external solution. Cav1.2 currents were elicited with voltage steps to 0mV every 20s. Activation of ORL-1 with nociceptin had no effect on Cav1.2 currents (n = 6). Data are expressed as mean ± SEM.

Article Snippet: The volume equivalent to 1-2mg of protein was then incubated with anti-Xpress monoclonal antibody (Invitrogen) or anti-Cav1.2 (CACNA1C) antibody (Alomone CAT# ACC-003) at a ratio of 1-2 μg antibody per 1mg of protein.

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Effect of ORL-1 on Cav1.2 calcium channels

doi: 10.64898/2026.07.03.736403

Figure Lengend Snippet: Effect of ORL1 on Cav1.2 trafficking. Cav1.2 membrane expression and endocytosis kinetics were explored by expressing a Cav1.2 containing an α-bungarotoxin binding site with or without ORL-1. Incubation with an α-bungarotoxin AF-594 conjugate at 17 °C for 30 minutes allowed for membrane-specific staining. Cells were then incubated in KRH at 37 °C for 0, 5, 10, 20 and 30 minutes before being fixed, permeabilized, and stained with an anti-α1c primary and anti-rabbit-488 conjugate. a) Visual representation of Cav1.2-BBS endocytosis experiments. At T0 (left), there is strong Cav1.2 membrane expression (red) and strong total Cav1.2 expression (green). At T10 (middle), there is a visible decrease in Cav1.2 membrane expression. At T30 (right), Cav1.2 membrane staining has drastically decreased. b) Cav1.2 membrane expression at T0, T5, T10, T20 and T30 was normalized to T0 for both Cav1.2 and Cav1.2+ORL-1 and then graphed as relative expression over time and fitted with a one-phase decay equation. ORL-1 did not alter Cav1.2 endocytosis kinetics (n = 5). c) Cav1.2 membrane expression is normalized to Cav1.2 total expression, then normalized to the Cav1.2 control condition as 1. The bar graph produced shows that co-expressing Cav1.2 with ORL-1 significantly decreases channel membrane expression compared to Cav1.2 alone (p < 0.0001, n = 220, n = 186). Data are expressed as mean ± SEM.

Article Snippet: The volume equivalent to 1-2mg of protein was then incubated with anti-Xpress monoclonal antibody (Invitrogen) or anti-Cav1.2 (CACNA1C) antibody (Alomone CAT# ACC-003) at a ratio of 1-2 μg antibody per 1mg of protein.

Techniques: Membrane, Expressing, Binding Assay, Incubation, Staining, Control, Produced, Immunofluorescence

Journal: bioRxiv

Article Title: Effect of ORL-1 on Cav1.2 calcium channels

doi: 10.64898/2026.07.03.736403

Figure Lengend Snippet: ORL-1 disrupts Cav1.2 forward trafficking. TsA-201 cells expressing Cav1.2-BBS with or without ORL-1 were incubated with an unlabelled α-bungarotoxin at 17 °C for 30 minutes to saturate membrane BBS binding sites before being incubated at 37 °C with α-bungarotoxin AF-594 for 0, 5, 10, 20 and 30 minutes. Only newly inserted membrane Cav1.2 channels during this time were therefore labelled with AF594. a) Visual representation of the forward trafficking experiments. At T0 (left), Cav1.2 membrane expression (red) is virtually nonexistent while total Cav1.2 expression (green) is strong. At T10 (middle), membrane staining has increased relative to T0 but is still minimal. At T30 (right), membrane expression has increased drastically compared to T0 and T10. b) Cav1.2 membrane expression in each cell, normalized to its own total expression for every time point. ORL-1 decreases Cav1.2 membrane expression starting at T5, and with the largest difference observed at T20 and T30. T0 (p < 0.0001, n = 87, n = 104), T10 (p = 0.0364, n = 122, n = 112, T20 p < 0.0001, n = 159, n = 113, T30 p < 0.0001, n = 171, n = 137. c) Cav1.2 membrane expression was normalized by total expression, then normalized to Cav1.2 control for each experiment. ORL-1 significantly decreases Cav1.2 membrane expression (p < 0.0001, n = 171, n = 137). Data are expressed as mean ± SEM.

Article Snippet: The volume equivalent to 1-2mg of protein was then incubated with anti-Xpress monoclonal antibody (Invitrogen) or anti-Cav1.2 (CACNA1C) antibody (Alomone CAT# ACC-003) at a ratio of 1-2 μg antibody per 1mg of protein.

Techniques: Expressing, Incubation, Membrane, Binding Assay, Staining, Control

Journal: International Journal of Molecular Sciences

Article Title: Super-Resolution 3D Imaging Reveals Disarray of Dyadic Calcium Ion Channels in Failing Hearts Expressing Low Thyroid Hormone Function

doi: 10.3390/ijms27125601

Figure Lengend Snippet: 3D STORM imaging and analysis of ventricular myocytes. Panels ( A ) Sham, ( B ) MI, and ( C ) MI+T3 show equivalent regions of interest (ROIs) of representative cells from each study group. Localizations are individual arbitrarily color-coded dots that represent signals in the fluorescent channels corresponding to immunolabeling of RyR2 (green), Ca V 1.2 (magenta) and Jph2 (red) proteins. Cell images after analysis of localizations in panels ( A – C ) are shown in respective panels ( a – c ). Cluster analysis of RyR2 signals by DBSCAN (density-based spatial clustering of applications with noise) algorithm shows RyR2 localizations connected by green mesh, and with each cluster encircled by a sphere (blue lattice) with a radius of 400 nm from the RyR2 cluster centroid. Magnification of the image in each panel is the same with xy-axes in the plane of the image, and the z-axis is perpendicular to the image. Scale bar 2 µm applies to all panels.

Article Snippet: CMs were first incubated in 0.02% TX-100/5% goat serum with anti-Ca V 1.2 (CACNA1C) rabbit polyclonal antibodies (1:400; ACC-003; Alomone Labs, Jerusalem, Israel) overnight at 4 °C, followed the next day by goat anti-rabbit IgG(H+L) DyLight 488 (1:2000; ab96883; Abcam, Cambridge, MA, USA) for 1 h at room temperature.

Techniques: Imaging, Analysis, Immunolabeling, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Super-Resolution 3D Imaging Reveals Disarray of Dyadic Calcium Ion Channels in Failing Hearts Expressing Low Thyroid Hormone Function

doi: 10.3390/ijms27125601

Figure Lengend Snippet: 3D STORM images of individual RyR2 clusters. Higher-magnification images of RyR2 clusters from Sham, MI, and MI+T3 cardiomyocytes as shown in a–c. RyR2 localizations forming single clusters were identified by DBSCAN algorithm and shown as green dots joined by green mesh. Localizations associated with Ca V 1.2 (magenta dots) and Jph2 (red dots) were counted within a sphere (lattice) measuring 400 nm radius (R) around the centroid of each RyR2 cluster. Not all localizations reside within the cluster-centered spheres. Reconstruction parameters, imaging conditions and cluster analysis constraints used were the same for images of all sample groups.

Article Snippet: CMs were first incubated in 0.02% TX-100/5% goat serum with anti-Ca V 1.2 (CACNA1C) rabbit polyclonal antibodies (1:400; ACC-003; Alomone Labs, Jerusalem, Israel) overnight at 4 °C, followed the next day by goat anti-rabbit IgG(H+L) DyLight 488 (1:2000; ab96883; Abcam, Cambridge, MA, USA) for 1 h at room temperature.

Techniques: Imaging, Analysis

Journal: International Journal of Molecular Sciences

Article Title: Super-Resolution 3D Imaging Reveals Disarray of Dyadic Calcium Ion Channels in Failing Hearts Expressing Low Thyroid Hormone Function

doi: 10.3390/ijms27125601

Figure Lengend Snippet: 3D STORM image analysis of total RyR2, Jph2 and Ca V 1.2 per cell. Cell volumes or ROI in 3D longitudinal images of cardiomyocytes measured 3 × 10 11 nm 3 . Violin plots show the distribution of individual cell results with each group median and quartiles as described in the legend to . Individual localization values were averaged from two ROIs of 10–12 cells/heart from 6–7 animals/group. The number of RyR2 clusters per ROI ( B ) and total numbers of RyR2, Jph2 and Ca V 1.2 localizations (locs) per ROI ( A , C , D ) were compared among the three study groups (Sham, MI, MI+T3) using nested one-way ANOVA and Tukey’s test for multigroup comparisons, * p < 0.05, τ p = 0.06. Violin plots show the distribution of results of individual cardiomyocytes (grey dots) isolated from 4 to 6 hearts per study group, with the group median (purple line) and quartiles (black lines) shown. The black triangles are the mean value of cells from each heart.

Article Snippet: CMs were first incubated in 0.02% TX-100/5% goat serum with anti-Ca V 1.2 (CACNA1C) rabbit polyclonal antibodies (1:400; ACC-003; Alomone Labs, Jerusalem, Israel) overnight at 4 °C, followed the next day by goat anti-rabbit IgG(H+L) DyLight 488 (1:2000; ab96883; Abcam, Cambridge, MA, USA) for 1 h at room temperature.

Techniques: Analysis, Isolation

Journal: International Journal of Molecular Sciences

Article Title: Super-Resolution 3D Imaging Reveals Disarray of Dyadic Calcium Ion Channels in Failing Hearts Expressing Low Thyroid Hormone Function

doi: 10.3390/ijms27125601

Figure Lengend Snippet: Analysis of RyR2 clusters and associated Jph2 and Ca V 1.2 in 3D STORM images. The violin plots in each analysis show the distribution of individual cell results (grey dots) with the mean value of each heart (black triangles) in the group and with the group median (purple line) and quartiles (black lines). All individual cell analyses were averaged from two equal ROIs per cell from 10 to 12 cells/heart of 6–7 animals per group. RyR2 clusters were identified by DBSCAN algorithm with subsequent measurements of cluster volume (nm 3 ) ( A ) and number of RyR2 localizations per cluster ( B ). Data in ( C , D ) are Jph2 and Ca V 1.2 localizations co-clustered within a sphere measured 400 nm radius of the RyR2 cluster centroid. Results in ( E – G ) represent percentage of RyR2 clusters without (w/o) co-clustered Jph2 or Ca V 1.2 or absent both proteins within the 400 nm sphere. Statistical analysis used nested one-way ANOVA with Tukey’s test for multiple group comparisons: significance at * p < 0.05. Violin plots show the distribution of results of individual cardiomyocytes (grey dots) isolated from 4 to 6 hearts per study group, with the group median (purple line) and quartiles (black lines) shown. The black triangles are the mean value of cells from each heart.

Article Snippet: CMs were first incubated in 0.02% TX-100/5% goat serum with anti-Ca V 1.2 (CACNA1C) rabbit polyclonal antibodies (1:400; ACC-003; Alomone Labs, Jerusalem, Israel) overnight at 4 °C, followed the next day by goat anti-rabbit IgG(H+L) DyLight 488 (1:2000; ab96883; Abcam, Cambridge, MA, USA) for 1 h at room temperature.

Techniques: Analysis, Isolation