ltcc α 1c  (Alomone Labs)


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    Alomone Labs ltcc α 1c
    Effects of Flu on the levels of <t>LTCC</t> <t>α</t> <t>1C</t> protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
    Ltcc α 1c, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    ltcc α 1c - by Bioz Stars, 2023-02
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    1) Product Images from "Fluvastatin Upregulates the α 1C Subunit of CaV1.2 Channel Expression in Vascular Smooth Muscle Cells via RhoA and ERK/p38 MAPK Pathways"

    Article Title: Fluvastatin Upregulates the α 1C Subunit of CaV1.2 Channel Expression in Vascular Smooth Muscle Cells via RhoA and ERK/p38 MAPK Pathways

    Journal: Disease Markers

    doi: 10.1155/2014/237067

    Effects of Flu on the levels of LTCC α 1C protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
    Figure Legend Snippet: Effects of Flu on the levels of LTCC α 1C protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Techniques Used: Western Blot, Expressing, Incubation, Derivative Assay

    Effects of MAPK or ROCK inhibition on LTCC α 1C expression after PDGF stimulation in VSMCs. LTCC α 1C protein expression was evaluated by western blot analysis. Data was described as means ± SEM from three experiments performed in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus PDGF stimulated cells. MAPK: mitogen activated protein kinase; ROCK: Rho associated protein kinase; LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
    Figure Legend Snippet: Effects of MAPK or ROCK inhibition on LTCC α 1C expression after PDGF stimulation in VSMCs. LTCC α 1C protein expression was evaluated by western blot analysis. Data was described as means ± SEM from three experiments performed in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus PDGF stimulated cells. MAPK: mitogen activated protein kinase; ROCK: Rho associated protein kinase; LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Techniques Used: Inhibition, Expressing, Western Blot, Derivative Assay

    lcc α 1c  (Alomone Labs)


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    Alomone Labs lcc α 1c
    Lcc α 1c, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    ltcc α 1c  (Alomone Labs)


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    Alomone Labs ltcc α 1c
    Effects of Flu on the levels of <t>LTCC</t> <t>α</t> <t>1C</t> protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
    Ltcc α 1c, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ltcc α 1c - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Fluvastatin Upregulates the α 1C Subunit of CaV1.2 Channel Expression in Vascular Smooth Muscle Cells via RhoA and ERK/p38 MAPK Pathways"

    Article Title: Fluvastatin Upregulates the α 1C Subunit of CaV1.2 Channel Expression in Vascular Smooth Muscle Cells via RhoA and ERK/p38 MAPK Pathways

    Journal: Disease Markers

    doi: 10.1155/2014/237067

    Effects of Flu on the levels of LTCC α 1C protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
    Figure Legend Snippet: Effects of Flu on the levels of LTCC α 1C protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Techniques Used: Western Blot, Expressing, Incubation, Derivative Assay

    Effects of MAPK or ROCK inhibition on LTCC α 1C expression after PDGF stimulation in VSMCs. LTCC α 1C protein expression was evaluated by western blot analysis. Data was described as means ± SEM from three experiments performed in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus PDGF stimulated cells. MAPK: mitogen activated protein kinase; ROCK: Rho associated protein kinase; LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
    Figure Legend Snippet: Effects of MAPK or ROCK inhibition on LTCC α 1C expression after PDGF stimulation in VSMCs. LTCC α 1C protein expression was evaluated by western blot analysis. Data was described as means ± SEM from three experiments performed in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus PDGF stimulated cells. MAPK: mitogen activated protein kinase; ROCK: Rho associated protein kinase; LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Techniques Used: Inhibition, Expressing, Western Blot, Derivative Assay

    α 1c  (Alomone Labs)


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    Alomone Labs α 1c
    α 1c, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    alpha 1c subunit vdcc  (Alomone Labs)


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    Alomone Labs alpha 1c subunit vdcc
    Representative blots are shown for <t>VDCC</t> (A). The amount of the <t>alpha-1c</t> subunit of VDCC was determined as the ratio of VDCC to β-actin (B). Results are expressed as the mean ± SEM (n = 6). ** P <0.01 vs each 8-week rat strain. Confocal measurement of VDCC (C) was performed in rat aortic segments (14 µm).
    Alpha 1c Subunit Vdcc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    alpha 1c subunit vdcc - by Bioz Stars, 2023-02
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    Images

    1) Product Images from "Attenuation of L-Type Ca 2+ Channel Expression and Vasomotor Response in the Aorta with Age in Both Wistar-Kyoto and Spontaneously Hypertensive Rats"

    Article Title: Attenuation of L-Type Ca 2+ Channel Expression and Vasomotor Response in the Aorta with Age in Both Wistar-Kyoto and Spontaneously Hypertensive Rats

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0088975

    Representative blots are shown for VDCC (A). The amount of the alpha-1c subunit of VDCC was determined as the ratio of VDCC to β-actin (B). Results are expressed as the mean ± SEM (n = 6). ** P <0.01 vs each 8-week rat strain. Confocal measurement of VDCC (C) was performed in rat aortic segments (14 µm).
    Figure Legend Snippet: Representative blots are shown for VDCC (A). The amount of the alpha-1c subunit of VDCC was determined as the ratio of VDCC to β-actin (B). Results are expressed as the mean ± SEM (n = 6). ** P <0.01 vs each 8-week rat strain. Confocal measurement of VDCC (C) was performed in rat aortic segments (14 µm).

    Techniques Used:

    α 1c alomone  (Alomone Labs)


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    Alomone Labs α 1c alomone
    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing <t>α</t> <t>1C</t> , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.
    α 1c Alomone, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits"

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    Journal: eLife

    doi: 10.7554/eLife.49253

    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.
    Figure Legend Snippet: ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.

    Techniques Used: Transfection, Flow Cytometry, Fluorescence, Expressing, Labeling, Patch Clamp

    ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.
    Figure Legend Snippet: ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.

    Techniques Used: Transfection, Flow Cytometry, Fluorescence, Labeling

    ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.
    Figure Legend Snippet: ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.

    Techniques Used: Expressing, Two Tailed Test

    ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.
    Figure Legend Snippet: ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.

    Techniques Used: Infection, Expressing

    ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.
    Figure Legend Snippet: ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.

    Techniques Used: Infection, Expressing, Immunostaining


    Figure Legend Snippet:

    Techniques Used: Recombinant, Clone Assay, Mutagenesis, Plasmid Preparation, Software

    anti α 1c  (Alomone Labs)


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    Alomone Labs anti α 1c
    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing <t>α</t> <t>1C</t> , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.
    Anti α 1c, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti α 1c - by Bioz Stars, 2023-02
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    1) Product Images from "A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits"

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    Journal: eLife

    doi: 10.7554/eLife.49253

    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.
    Figure Legend Snippet: ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.

    Techniques Used: Transfection, Flow Cytometry, Fluorescence, Expressing, Labeling, Patch Clamp

    ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.
    Figure Legend Snippet: ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.

    Techniques Used: Transfection, Flow Cytometry, Fluorescence, Labeling

    ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.
    Figure Legend Snippet: ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.

    Techniques Used: Expressing, Two Tailed Test

    ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.
    Figure Legend Snippet: ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.

    Techniques Used: Infection, Expressing

    ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.
    Figure Legend Snippet: ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.

    Techniques Used: Infection, Expressing, Immunostaining


    Figure Legend Snippet:

    Techniques Used: Recombinant, Clone Assay, Mutagenesis, Plasmid Preparation, Software

    alpha 1c subunit  (Alomone Labs)


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    anti α 1c polyclonal antibody  (Alomone Labs)


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    Effects of Flu on the levels of <t>LTCC</t> <t>α</t> <t>1C</t> protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.
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    Alomone Labs antibody anti α 1c polyclonal antibody
    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing <t>α</t> <t>1C</t> , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.
    Antibody Anti α 1c Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of Flu on the levels of LTCC α 1C protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Journal: Disease Markers

    Article Title: Fluvastatin Upregulates the α 1C Subunit of CaV1.2 Channel Expression in Vascular Smooth Muscle Cells via RhoA and ERK/p38 MAPK Pathways

    doi: 10.1155/2014/237067

    Figure Lengend Snippet: Effects of Flu on the levels of LTCC α 1C protein determined by western blot analysis. PDGF time-dependently inhibited LTCC α 1C expression, which could be prevented by fluvastatin. (a) Dose-effect response of PDGF stimulation on the LTCC α 1C expression in VSMCs. (b) Time-effect response of Flu on the LTCC α 1C expression incubated by PDGF. β-actin served as the loading control. Data was represented as means ± SE of 3 separate experiments conducted in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus cells incubation with PDGF. LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Article Snippet: Primary antibodies were used at the indicated dilutions as follows: LTCC α 1C , 1 : 200 (Alomone); β -actin, 1 : 1000; anti-p-p38 MAPK, 1 : 1000; anti-total-p38 MAPK, 1 : 1000; anti-p-JNK, 1 : 1000; anti-total-JNK 1 : 1000; anti-p-ERK1/2 (1 : 1000); and anti-total-ERK1/2 (1 : 1000); all were from Cell Signalling Technology (Danvers, MA).

    Techniques: Western Blot, Expressing, Incubation, Derivative Assay

    Effects of MAPK or ROCK inhibition on LTCC α 1C expression after PDGF stimulation in VSMCs. LTCC α 1C protein expression was evaluated by western blot analysis. Data was described as means ± SEM from three experiments performed in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus PDGF stimulated cells. MAPK: mitogen activated protein kinase; ROCK: Rho associated protein kinase; LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Journal: Disease Markers

    Article Title: Fluvastatin Upregulates the α 1C Subunit of CaV1.2 Channel Expression in Vascular Smooth Muscle Cells via RhoA and ERK/p38 MAPK Pathways

    doi: 10.1155/2014/237067

    Figure Lengend Snippet: Effects of MAPK or ROCK inhibition on LTCC α 1C expression after PDGF stimulation in VSMCs. LTCC α 1C protein expression was evaluated by western blot analysis. Data was described as means ± SEM from three experiments performed in triplicate. * P < 0.05 versus blank control and # P < 0.05 versus PDGF stimulated cells. MAPK: mitogen activated protein kinase; ROCK: Rho associated protein kinase; LTCC α 1C : L-type calcium channel α 1C subunit; Flu: fluvastatin; and PDGF: platelet derived growth factor.

    Article Snippet: Primary antibodies were used at the indicated dilutions as follows: LTCC α 1C , 1 : 200 (Alomone); β -actin, 1 : 1000; anti-p-p38 MAPK, 1 : 1000; anti-total-p38 MAPK, 1 : 1000; anti-p-JNK, 1 : 1000; anti-total-JNK 1 : 1000; anti-p-ERK1/2 (1 : 1000); and anti-total-ERK1/2 (1 : 1000); all were from Cell Signalling Technology (Danvers, MA).

    Techniques: Inhibition, Expressing, Western Blot, Derivative Assay

    Representative blots are shown for VDCC (A). The amount of the alpha-1c subunit of VDCC was determined as the ratio of VDCC to β-actin (B). Results are expressed as the mean ± SEM (n = 6). ** P <0.01 vs each 8-week rat strain. Confocal measurement of VDCC (C) was performed in rat aortic segments (14 µm).

    Journal: PLoS ONE

    Article Title: Attenuation of L-Type Ca 2+ Channel Expression and Vasomotor Response in the Aorta with Age in Both Wistar-Kyoto and Spontaneously Hypertensive Rats

    doi: 10.1371/journal.pone.0088975

    Figure Lengend Snippet: Representative blots are shown for VDCC (A). The amount of the alpha-1c subunit of VDCC was determined as the ratio of VDCC to β-actin (B). Results are expressed as the mean ± SEM (n = 6). ** P <0.01 vs each 8-week rat strain. Confocal measurement of VDCC (C) was performed in rat aortic segments (14 µm).

    Article Snippet: Rabbit anti-Cav1.2, the alpha-1c subunit VDCC, primary antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques:

    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.

    Article Snippet: Primary antibodies and working dilutions were as follows: α 1C : Alomone, 1:1000; UC Davis/NIH NeuroMab Facility, clone N263/31, 1:200.

    Techniques: Transfection, Flow Cytometry, Fluorescence, Expressing, Labeling, Patch Clamp

    ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.

    Article Snippet: Primary antibodies and working dilutions were as follows: α 1C : Alomone, 1:1000; UC Davis/NIH NeuroMab Facility, clone N263/31, 1:200.

    Techniques: Transfection, Flow Cytometry, Fluorescence, Labeling

    ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.

    Article Snippet: Primary antibodies and working dilutions were as follows: α 1C : Alomone, 1:1000; UC Davis/NIH NeuroMab Facility, clone N263/31, 1:200.

    Techniques: Expressing, Two Tailed Test

    ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.

    Article Snippet: Primary antibodies and working dilutions were as follows: α 1C : Alomone, 1:1000; UC Davis/NIH NeuroMab Facility, clone N263/31, 1:200.

    Techniques: Infection, Expressing

    ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.

    Article Snippet: Primary antibodies and working dilutions were as follows: α 1C : Alomone, 1:1000; UC Davis/NIH NeuroMab Facility, clone N263/31, 1:200.

    Techniques: Infection, Expressing, Immunostaining

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet:

    Article Snippet: Primary antibodies and working dilutions were as follows: α 1C : Alomone, 1:1000; UC Davis/NIH NeuroMab Facility, clone N263/31, 1:200.

    Techniques: Recombinant, Clone Assay, Mutagenesis, Plasmid Preparation, Software

    ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Cartoon of experimental strategy. BBS-α 1C was transfected in HEK293 cells with each YFP-Ca V β and either CFP or nb.F3-P2A-CFP. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , Ca V β 1 and CFP (left) or nb.F3 (right). ( c ) Cumulative distribution histogram of Alexa-647 (left) or YFP fluorescence (right) from CFP (black) or nb.F3 (red) expressing cells. YFP-positive cells (n > 5,000 cells/experiment) were selected for analysis; the threshold for 647 labeling and YFP fluorescence above background is represented with the dashed line. ( d ) Summary flow cytometry data of surface (647, filled) and total (YFP, patterned) levels of BBS-α 1C . Data from nb.F3 was normalized to CFP control group. N = 4 separate experiments, error bars, s.e.m. ( e ) population I-V curves from whole-cell patch clamp measurements in HEK293 cells expressing α 1C , Ca V β 2a , and CFP (black, n = 9) or nb.F3 (red, n = 12). Data are means ± s.e.m.

    Article Snippet: Antibody , Anti-α 1C , Alomone , Cat#: ACC-003 , 1:1000 WB/IF.

    Techniques: Transfection, Flow Cytometry, Fluorescence, Expressing, Labeling, Patch Clamp

    ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Schematic of experimental design. HEK293 cells were transfected with BBS-α 1C , YFP-Ca V β, and either nb.F3, nb.F3-Nedd4L or nb.F3-Nedd4L[C942S]. ( b ) Exemplar flow cytometry dot plot of cells transfected with BBS-α 1C , YFP-Ca V β 1b and nb.F3 (left), nb.F3-Nedd4L (middle) or nb.F3-Nedd4L[C942S] (right). ( c,d ) Histogram of YFP ( c ) and Alexa-647 ( d ) fluorescence from samples in ( b ) (left) and summary data from N = 3 separate experiments (right). YFP-positive cells (n > 5,000 cells per experiment) were selected for analysis, the threshold for 647 labeling above background is represented with the dashed line. *p<0.05 compared with control, one-way ANOVA with Tukey’s multiple comparison test.

    Article Snippet: Antibody , Anti-α 1C , Alomone , Cat#: ACC-003 , 1:1000 WB/IF.

    Techniques: Transfection, Flow Cytometry, Fluorescence, Labeling

    ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Population I-V curves from HEK293 expressing α 1C + β 1b + α 2 δ−1 with either nb.F3 (black, I peak, 0mV = −48.4 ± 8.4 pA/pF, n = 12) or Ca V -aβlator (red, I peak, 0mV = −0.93 ± 0.16 pA/pF, n = 8). ( b-d ) Same format as ( a ) for cells expressing reconstituted Ca V 1.3 ( b ), Ca V 2.1 ( c ), or Ca V 2.3 ( d ) channels. Data are means ± s.e.m.†p<0.01 compared with control, unpaired, two-tailed Student’s t-test.

    Article Snippet: Antibody , Anti-α 1C , Alomone , Cat#: ACC-003 , 1:1000 WB/IF.

    Techniques: Expressing, Two Tailed Test

    ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Confocal images (top) and exemplar traces from whole-cell recordings of uninfected guinea pig cardiomyocytes (left), or infected with adenovirus expressing either Ca V -βlator (middle) or nb.F3-Nedd4L[C942S] (right). Scale bar 0.2nA, 10 ms. ( b ) Population I-V curves from cardiomyocytes expressing Ca V -βlator (red), nb.F3-Nedd4L[C942S] (green), or an uninfected control (black). ( c ) Left, exemplar confocal images of cardiomyocytes fixed and immunostained with α 1C (green) and ryanodine receptor (RyR2, magenta) antibodies. Yellow box indicates region of high-zoom merge image.Right, co-localization between α 1C and RyR in uninfected cardiomyocytes (gray, PCC = 0.47 ± 0.02, n = 15), and those expressing either Ca V -aβlator (red, PCC = 0.24 ± 0.02 n = 19), or nb.F3-Nedd4L[C942S] (green, PCC = 0.50 ± 0.01, n = 17). ( d ) Left, exemplar confocal images of fixed cardiomyocytes immunostained with α 1C (green) and Rab7 (magenta) antibodies. Yellow box indicates region of high-zoom merge image. Right, colocalization between α 1C and Rab7 in uninfected cardiomyocytes (gray, PCC = 0.29 ± 0.02, n = 16), and those expressing either Ca V -aβlator (red, PCC = 0.42 ± 0.02, n = 18), or nb.F3-Nedd4L[C942S] (green, PCC = 0.30 ± 0.03, n = 16). ( e ) Pulldown of α 1C in HEK293 cells expressing α 1C , β 1b and either CFP, nb.F3, Ca V -aβlator, or nb.F3-Nedd4L-[C942S]. Top, probing pulldown with α 1C antibody. Bottom, same blot stripped and re-probed with ubiquitin antibody. ( f ) Quantification of four separate experiments, as performed in ( e ). Data are means ± s.e.m for each point. *p<0.05 compared to control, one-way ANOVA with Tukey’s multiple comparison test. ( g ) Pulldown of Ca V β 1b , as in ( e ). Left, probing with Ca V β 1b . Right, same blot stripped and re-probed with ubiquitin antibody. ( h ) Cartoon illustrating Ca V -aβlator-induced relocation of Ca V 1.2 from dyads to Rab7-positive late endosomes in cardiomyocytes.

    Article Snippet: Antibody , Anti-α 1C , Alomone , Cat#: ACC-003 , 1:1000 WB/IF.

    Techniques: Infection, Expressing

    ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet: ( a ) Comparison of total α 1C levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 19). ( b ). Comparison of total β 2 levels in uninfected cardiomyocytes (gray, n = 15) and those infected with nb.F3-Nedd4L (red, n = 16), normalized to uninfected controls. Data are means ± s.e.m. ( c ) Left, exemplar confocal images of uninfected (top) or F3-Nedd4L-infected (bottom) guinea pig cardiomyocytes, fixed and immunostained with antibodies towards α 1C (left) and LAMP1 (middle). Right, colocalization between α 1C and Rab5 in uninfected cardiomyocytes (gray, n = 7) and those expressing F3-Nedd4L (red, n = 7). Data are means ± s.e.m. ( d ) as in ( c ), showing immunostaining and colocalization analysis of cardiomyocytes immunostained against α 1C and Rab5.

    Article Snippet: Antibody , Anti-α 1C , Alomone , Cat#: ACC-003 , 1:1000 WB/IF.

    Techniques: Infection, Expressing, Immunostaining

    Journal: eLife

    Article Title: A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

    doi: 10.7554/eLife.49253

    Figure Lengend Snippet:

    Article Snippet: Antibody , Anti-α 1C , Alomone , Cat#: ACC-003 , 1:1000 WB/IF.

    Techniques: Recombinant, Clone Assay, Mutagenesis, Plasmid Preparation, Software