cav1 2 α1c  (Alomone Labs)

Journal: International Journal of Molecular Sciences

Article Title: Thyroid Hormone Upregulates Cav1.2 Channels in Cardiac Cells via the Downregulation of the Channels’ β4 Subunit

doi: 10.3390/ijms251910798

Figure Lengend Snippet: Relative mRNA expression and α1c protein abundance in H9c2 cells after incubation with T3. ( A , B ) Means ± SEMs of relative α1c subunit mRNA expression and protein abundance in H9c2 cells from three separate experiments. Insets show representative blots under control conditions (lanes a, c, e, and g) and 2, 6, 24, and 48 h after T3 treatment (lanes b, d, f, and h, respectively). Tubulin bands were used to normalize α1c subunit density values. In panels ( A , B ) each symbol represents a separate experiment. Original blots are presented in . * p < 0.05. ** p < 0.01.

Article Snippet: The antibody sources were polyclonal anti-rabbit α1c (ACC-003, 1:200; Alomone Labs, Jerusalem, Israel) and polyclonal anti-rabbit CACNB4 (1:500, 1:1000, A9304; ABclonal Technology, Woburn, MA, USA).

Techniques: Expressing, Incubation, Control

Journal: International Journal of Molecular Sciences

Article Title: Thyroid Hormone Upregulates Cav1.2 Channels in Cardiac Cells via the Downregulation of the Channels’ β4 Subunit

doi: 10.3390/ijms251910798

Figure Lengend Snippet: Relative α1c mRNA expression and protein abundance in H9c2 cells after Cavβ4 subunit overexpression. ( A ) Means ± SEMs of relative Cavβ4 protein abundance under control conditions and after Cavβ4 overexpression ( n = 3). The inset shows representative blots of the Cavβ4 subunit and GAPDH bands; the latter was used to normalize Cavβ4 subunit density values. Original blots are presented in . ( B ) Means ± SEMs of relative α1c subunit mRNA expression from control and Cavβ4 overexpression experiments ( n = 14). ( C ) Means ± SEMs of α1c protein abundance from control and Cavβ4 overexpression experiments ( n = 10). The inset shows representative blots of α1c subunit and tubulin bands under control conditions (lanes a and c) and after Cavβ4 overexpression (lanes b and d) from two separate experiments. Original blots are presented in . In panels ( A , B , C ), each symbol represents a separate experiment. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The antibody sources were polyclonal anti-rabbit α1c (ACC-003, 1:200; Alomone Labs, Jerusalem, Israel) and polyclonal anti-rabbit CACNB4 (1:500, 1:1000, A9304; ABclonal Technology, Woburn, MA, USA).

Techniques: Expressing, Over Expression, Control

Journal: International Journal of Molecular Sciences

Article Title: Thyroid Hormone Upregulates Cav1.2 Channels in Cardiac Cells via the Downregulation of the Channels’ β4 Subunit

doi: 10.3390/ijms251910798

Figure Lengend Snippet: Role of Cavβ4 in pCREB regulation by T3. Schematic showing signaling mechanisms hypothetically involved in thyroid hormone actions on the CACNA1C gene via pCREB and the release of Cavβ4 inhibition of α1c subunit expression in ventricular cardiac cells.

Article Snippet: The antibody sources were polyclonal anti-rabbit α1c (ACC-003, 1:200; Alomone Labs, Jerusalem, Israel) and polyclonal anti-rabbit CACNB4 (1:500, 1:1000, A9304; ABclonal Technology, Woburn, MA, USA).

Techniques: Inhibition, Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Palmitoylation of the pore-forming subunit of Ca(v)1.2 controls channel voltage sensitivity and calcium transients in cardiac myocytes

doi: 10.1073/pnas.2207887120

Figure Lengend Snippet: Ca(v) 1.2 α1C subunit is palmitoylated in mouse, rabbit, and human ventricular tissues. Palmitoylated proteins were purified by resin-assisted capture of acylated proteins (acyl-RAC) and immunoblotted as shown. The bar chart below each blot indicates the abundance of Ca(v)1.2 α1C and caveolin 3 (Cav3) in the purified palmitoylated fraction (Palm) relative to the corresponding unfractionated lysate (UF). N = 4 (mouse), N = 8 (rabbit), N = 7 (human). *** P < 0.001, unpaired t test. Error bars represent SEM.

Article Snippet: Anti-Ca(v)1.2 α1C subunit antibodies raised in rabbit and guinea pig were obtained from Alomone Labs, antibodies against flotillin 2 and caveolin 3 from BD Biosciences, and anti-GFP antibodies from Abcam and Protein Tech.

Techniques: Purification

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Palmitoylation of the pore-forming subunit of Ca(v)1.2 controls channel voltage sensitivity and calcium transients in cardiac myocytes

doi: 10.1073/pnas.2207887120

Figure Lengend Snippet: Palmitoylation site conservation in human Ca(v)1 channel isoforms. For clarity, regions of the rabbit α1C splice variant of Ca(v)1.2 CACH2A (UniProt accession number P15381) with palmitoylated cysteines identified in this investigation numbered and highlighted in red are shown above the corresponding regions of the human channels. Numbers at the end of each sequence are the numbering of the final amino acid in the region of each Ca(v)1 isoform shown. Human Ca(v)1 isoforms (UniProt accession numbers shown) were aligned using Clustal Ω. “*” below an amino acid indicates 100% conservation between isoforms; “:” indicates amino acids of highly similar properties; “.” indicates amino acids of weakly similar properties. The palmitoylation site in the Ca(v)1.2 N terminus is conserved in all isoforms. Ca(v)1.1 does not possess a cysteine analogous to C519 in the I–II linker, but Ca(v)1.3 and 1.4 do. C543 is unique to Ca(v)1.2.

Article Snippet: Anti-Ca(v)1.2 α1C subunit antibodies raised in rabbit and guinea pig were obtained from Alomone Labs, antibodies against flotillin 2 and caveolin 3 from BD Biosciences, and anti-GFP antibodies from Abcam and Protein Tech.

Techniques: Variant Assay, Sequencing

Journal: PLoS ONE

Article Title: Differential Calcium Signaling Mediated by Voltage-Gated Calcium Channels in Rat Retinal Ganglion Cells and Their Unmyelinated Axons

doi: 10.1371/journal.pone.0084507

Figure Lengend Snippet: Ca channel α 1 subunits co-express with the ganglion cell antibody RBPMS in vertical sections showing the inner plexiform layer (IPL), the ganglion cell layer (GCL) and the nerve fiber layer (NFL). ( Top row ) Double immunostaining with α1C subunit and RBPMS antibodies shows colocalization in the RGC somata (filled arrow). RGC axons (arrowhead) as well as putative Müller and neuronal cell processes in the IPL showed immunostaining. Control preadsorption of α1C antibody with the immunization peptide showed no staining. ( Second row ) Double immunostaining with α1D subunit and RBPMS antibodies shows colocalization in the RGC somata (filled arrow). Numerous processes in the IPL as well as the RGC axons throughout the NFL (arrowhead) were also labelled. Control preadsorption of α1D antibody with the immunization peptide showed no staining. ( Third row ) Double immunostaining with α1B subunit and RBPMS antibodies shows colocalization in RGC somata (filled arrow). Processes in the IPL, putative displaced amacrine cells in the GCL (open arrows) and RGC axons throughout the NFL (arrowheads) showed immunostaining. Control preadsorption of α1B antibody with the immunization peptide showed no staining. ( Bottom row ) Double immunostaining with α1A subunit and RBPMS antibodies shows colocalization in the RGC somata (filled arrow). Labelling in the IPL and in putative displaced amacrine cells in the GCL (open arrows) was also seen. Control preadsorption of α1A antibody with the immunization peptide showed no staining. Scale bar is 20 µm.

Article Snippet: Preadsorption controls were also performed with the α1C and α1B antibodies (Alomone; ACC-003 and ACC-002, respectively) at a final concentration of 1 µg/ml for 12–16 h overnight at 4°C.

Techniques: Double Immunostaining, Immunostaining, Staining

Journal: PLoS ONE

Article Title: Differential Calcium Signaling Mediated by Voltage-Gated Calcium Channels in Rat Retinal Ganglion Cells and Their Unmyelinated Axons

doi: 10.1371/journal.pone.0084507

Figure Lengend Snippet: ( Top row ) Rat wholemount retina triple labelled with RBPMS (red), α1C VGCC subunit (green), and NF-M (white) antibodies. α1C colocalized with RBPMS, which labels RGC somata (filled arrow), and NF-M, which labels RGC axons (arrowhead), as well as putative Müller cell endfeet. Stack of five z-axis optical sections each of 0.3 µm thickness. ( Lower row ) Rat wholemount retina labelled with RBPMS (red), α1D VGCC subunit (green), and NF-M (white) antibodies. α1D was localized to RGC somata (RBPMS) (filled arrow), RGC axons (NF-M) (arrowhead) and putative displaced amacrine cells (open arrow). Stack of four optical sections each of 0.3 µm thickness. Scale bar is 20 µm.

Article Snippet: Preadsorption controls were also performed with the α1C and α1B antibodies (Alomone; ACC-003 and ACC-002, respectively) at a final concentration of 1 µg/ml for 12–16 h overnight at 4°C.

Techniques: