anti cav1 2 rabbit polyclonal  (Alomone Labs)


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    Structured Review

    Alomone Labs anti cav1 2 rabbit polyclonal
    <t>CaV1.2</t> downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P
    Anti Cav1 2 Rabbit Polyclonal, 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
    https://www.bioz.com/result/anti cav1 2 rabbit polyclonal/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 rabbit polyclonal - by Bioz Stars, 2022-01
    86/100 stars

    Images

    1) Product Images from "TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets"

    Article Title: TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI124481

    CaV1.2 downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P
    Figure Legend Snippet: CaV1.2 downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P

    Techniques Used: Expressing, Transfection, Plasmid Preparation, In Situ Hybridization

    Loss of TDP-43 reduces insulin secretion by downregulating CaV1.2 calcium channel expression in MIN6 cells.
    Figure Legend Snippet: Loss of TDP-43 reduces insulin secretion by downregulating CaV1.2 calcium channel expression in MIN6 cells.

    Techniques Used: Expressing

    TDP-43 regulates the transcription of CaV1.2 calcium channels. ( A ) Immunoprecipitation (IP) of lysates from MIN6 cells overexpressing V5-tagged TDP-43 with rabbit IgG or an anti-V5 antibody (IP-V5), analyzed with immunoblotting with the indicated antibody. ( B ) RNA-IP of V5 in cultured MIN6 cells. Bound RNA was analyzed with qRT-PCR using the indicated primers. IP efficiency was calculated relative to input. Representative data from triplicate experiments are shown. ( C ) Schematics of mature mRNA (exon), premature mRNA (intron), and putative promoter are shown over the Cacna1c gene. ( D ) Each mature mRNA (exon) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( E ) mRNA stability was measured in MIN6 cells (control, T1) treated with 10 mg/mL actinomycin D with qRT-PCR using the indicated Cacna1c and Tuba1a primers ( n = 4 each, 2-way ANOVA). The mRNA levels relative to pretreatment were plotted against time after treatment. Residual mRNA levels were compared at 2, 4, 8, and 12 hours after treatment. ( F ) Each premature mRNA (intron) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( G ) Luciferase reporter assay of the –1542/+257 mouse Cacna1c promoter ( n = 10 each, unpaired t test). ( H ) Human CACNA1C promoter luciferase reporter assay using the LightSwitch promoter reporter GoClone collection ( n = 11 each, unpaired t test). Values are mean ± SEM. * P
    Figure Legend Snippet: TDP-43 regulates the transcription of CaV1.2 calcium channels. ( A ) Immunoprecipitation (IP) of lysates from MIN6 cells overexpressing V5-tagged TDP-43 with rabbit IgG or an anti-V5 antibody (IP-V5), analyzed with immunoblotting with the indicated antibody. ( B ) RNA-IP of V5 in cultured MIN6 cells. Bound RNA was analyzed with qRT-PCR using the indicated primers. IP efficiency was calculated relative to input. Representative data from triplicate experiments are shown. ( C ) Schematics of mature mRNA (exon), premature mRNA (intron), and putative promoter are shown over the Cacna1c gene. ( D ) Each mature mRNA (exon) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( E ) mRNA stability was measured in MIN6 cells (control, T1) treated with 10 mg/mL actinomycin D with qRT-PCR using the indicated Cacna1c and Tuba1a primers ( n = 4 each, 2-way ANOVA). The mRNA levels relative to pretreatment were plotted against time after treatment. Residual mRNA levels were compared at 2, 4, 8, and 12 hours after treatment. ( F ) Each premature mRNA (intron) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( G ) Luciferase reporter assay of the –1542/+257 mouse Cacna1c promoter ( n = 10 each, unpaired t test). ( H ) Human CACNA1C promoter luciferase reporter assay using the LightSwitch promoter reporter GoClone collection ( n = 11 each, unpaired t test). Values are mean ± SEM. * P

    Techniques Used: Immunoprecipitation, Cell Culture, Quantitative RT-PCR, Luciferase, Reporter Assay

    2) Product Images from "TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets"

    Article Title: TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI124481

    CaV1.2 downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P
    Figure Legend Snippet: CaV1.2 downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P

    Techniques Used: Expressing, Transfection, Plasmid Preparation, In Situ Hybridization

    Loss of TDP-43 reduces insulin secretion by downregulating CaV1.2 calcium channel expression in MIN6 cells.
    Figure Legend Snippet: Loss of TDP-43 reduces insulin secretion by downregulating CaV1.2 calcium channel expression in MIN6 cells.

    Techniques Used: Expressing

    TDP-43 regulates the transcription of CaV1.2 calcium channels. ( A ) Immunoprecipitation (IP) of lysates from MIN6 cells overexpressing V5-tagged TDP-43 with rabbit IgG or an anti-V5 antibody (IP-V5), analyzed with immunoblotting with the indicated antibody. ( B ) RNA-IP of V5 in cultured MIN6 cells. Bound RNA was analyzed with qRT-PCR using the indicated primers. IP efficiency was calculated relative to input. Representative data from triplicate experiments are shown. ( C ) Schematics of mature mRNA (exon), premature mRNA (intron), and putative promoter are shown over the Cacna1c gene. ( D ) Each mature mRNA (exon) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( E ) mRNA stability was measured in MIN6 cells (control, T1) treated with 10 mg/mL actinomycin D with qRT-PCR using the indicated Cacna1c and Tuba1a primers ( n = 4 each, 2-way ANOVA). The mRNA levels relative to pretreatment were plotted against time after treatment. Residual mRNA levels were compared at 2, 4, 8, and 12 hours after treatment. ( F ) Each premature mRNA (intron) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( G ) Luciferase reporter assay of the –1542/+257 mouse Cacna1c promoter ( n = 10 each, unpaired t test). ( H ) Human CACNA1C promoter luciferase reporter assay using the LightSwitch promoter reporter GoClone collection ( n = 11 each, unpaired t test). Values are mean ± SEM. * P
    Figure Legend Snippet: TDP-43 regulates the transcription of CaV1.2 calcium channels. ( A ) Immunoprecipitation (IP) of lysates from MIN6 cells overexpressing V5-tagged TDP-43 with rabbit IgG or an anti-V5 antibody (IP-V5), analyzed with immunoblotting with the indicated antibody. ( B ) RNA-IP of V5 in cultured MIN6 cells. Bound RNA was analyzed with qRT-PCR using the indicated primers. IP efficiency was calculated relative to input. Representative data from triplicate experiments are shown. ( C ) Schematics of mature mRNA (exon), premature mRNA (intron), and putative promoter are shown over the Cacna1c gene. ( D ) Each mature mRNA (exon) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( E ) mRNA stability was measured in MIN6 cells (control, T1) treated with 10 mg/mL actinomycin D with qRT-PCR using the indicated Cacna1c and Tuba1a primers ( n = 4 each, 2-way ANOVA). The mRNA levels relative to pretreatment were plotted against time after treatment. Residual mRNA levels were compared at 2, 4, 8, and 12 hours after treatment. ( F ) Each premature mRNA (intron) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( G ) Luciferase reporter assay of the –1542/+257 mouse Cacna1c promoter ( n = 10 each, unpaired t test). ( H ) Human CACNA1C promoter luciferase reporter assay using the LightSwitch promoter reporter GoClone collection ( n = 11 each, unpaired t test). Values are mean ± SEM. * P

    Techniques Used: Immunoprecipitation, Cell Culture, Quantitative RT-PCR, Luciferase, Reporter Assay

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    Alomone Labs cav1 2
    <t>Cav1.2</t> knock-down prevents astrocyte activation after scratch
    Cav1 2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cav1 2/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 - by Bioz Stars, 2022-01
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    95
    Alomone Labs acc 003 antibody
    Location of antibody epitopes. Shown is a schematic of the Ca v 1.2 α 1 1.2 subunit, in which regions used as immunogens for the depicted antibodies are identified by arrows. Exact residues are listed in the table and numbered according to α 1 1.2 given in Gene Bank Accession number CAA33546. FP1, CNC1, and <t>ACC-003</t> are directed against the loop between domains II and III, pS1700 against phosphorylated S1700, pS1928 against phosphorylated S1928, and CNC2 against residues 2122-2138 of α 1 1.2, which are ~40 residues upstream of the very C terminus of α 1 1.2.
    Acc 003 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/acc 003 antibody/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    acc 003 antibody - by Bioz Stars, 2022-01
    95/100 stars
      Buy from Supplier

    86
    Alomone Labs anti cav1 2 rabbit polyclonal
    <t>CaV1.2</t> downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P
    Anti Cav1 2 Rabbit Polyclonal, 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
    https://www.bioz.com/result/anti cav1 2 rabbit polyclonal/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 rabbit polyclonal - by Bioz Stars, 2022-01
    86/100 stars
      Buy from Supplier

    Image Search Results


    Cav1.2 knock-down prevents astrocyte activation after scratch

    Journal: Glia

    Article Title: L-type voltage-operated calcium channels contribute to astrocyte activation in vitro

    doi: 10.1002/glia.23013

    Figure Lengend Snippet: Cav1.2 knock-down prevents astrocyte activation after scratch

    Article Snippet: In contrast, RT-PCR, western blot and immunocytochemistry experiments show that Cav1.2 and Cav1.3 are highly expressed in cortical astrocytes ( ). shows representative images of labeled cultured astrocytes at 6 days in vitro ( DIV ); while GFAP abundantly labeled cell processes and cell bodies, labeling with Cav1.2 and Cav1.3 antibodies displayed a punctate staining on the cells surface ( ).

    Techniques: Activation Assay

    LPS enhance the activity and the expression of Cav1.2 Ca ++ channels in astrocytes

    Journal: Glia

    Article Title: L-type voltage-operated calcium channels contribute to astrocyte activation in vitro

    doi: 10.1002/glia.23013

    Figure Lengend Snippet: LPS enhance the activity and the expression of Cav1.2 Ca ++ channels in astrocytes

    Article Snippet: In contrast, RT-PCR, western blot and immunocytochemistry experiments show that Cav1.2 and Cav1.3 are highly expressed in cortical astrocytes ( ). shows representative images of labeled cultured astrocytes at 6 days in vitro ( DIV ); while GFAP abundantly labeled cell processes and cell bodies, labeling with Cav1.2 and Cav1.3 antibodies displayed a punctate staining on the cells surface ( ).

    Techniques: Activity Assay, Expressing

    Cav1.2 KO astrocytes are not sensitive to LPS

    Journal: Glia

    Article Title: L-type voltage-operated calcium channels contribute to astrocyte activation in vitro

    doi: 10.1002/glia.23013

    Figure Lengend Snippet: Cav1.2 KO astrocytes are not sensitive to LPS

    Article Snippet: In contrast, RT-PCR, western blot and immunocytochemistry experiments show that Cav1.2 and Cav1.3 are highly expressed in cortical astrocytes ( ). shows representative images of labeled cultured astrocytes at 6 days in vitro ( DIV ); while GFAP abundantly labeled cell processes and cell bodies, labeling with Cav1.2 and Cav1.3 antibodies displayed a punctate staining on the cells surface ( ).

    Techniques:

    Cav1.2 knock-down prevents astrocyte activation by LPS

    Journal: Glia

    Article Title: L-type voltage-operated calcium channels contribute to astrocyte activation in vitro

    doi: 10.1002/glia.23013

    Figure Lengend Snippet: Cav1.2 knock-down prevents astrocyte activation by LPS

    Article Snippet: In contrast, RT-PCR, western blot and immunocytochemistry experiments show that Cav1.2 and Cav1.3 are highly expressed in cortical astrocytes ( ). shows representative images of labeled cultured astrocytes at 6 days in vitro ( DIV ); while GFAP abundantly labeled cell processes and cell bodies, labeling with Cav1.2 and Cav1.3 antibodies displayed a punctate staining on the cells surface ( ).

    Techniques: Activation Assay

    Location of antibody epitopes. Shown is a schematic of the Ca v 1.2 α 1 1.2 subunit, in which regions used as immunogens for the depicted antibodies are identified by arrows. Exact residues are listed in the table and numbered according to α 1 1.2 given in Gene Bank Accession number CAA33546. FP1, CNC1, and ACC-003 are directed against the loop between domains II and III, pS1700 against phosphorylated S1700, pS1928 against phosphorylated S1928, and CNC2 against residues 2122-2138 of α 1 1.2, which are ~40 residues upstream of the very C terminus of α 1 1.2.

    Journal: F1000Research

    Article Title: Proteolytic processing of the L-type Ca2+ channel alpha11.2 subunit in neurons

    doi: 10.12688/f1000research.11808.1

    Figure Lengend Snippet: Location of antibody epitopes. Shown is a schematic of the Ca v 1.2 α 1 1.2 subunit, in which regions used as immunogens for the depicted antibodies are identified by arrows. Exact residues are listed in the table and numbered according to α 1 1.2 given in Gene Bank Accession number CAA33546. FP1, CNC1, and ACC-003 are directed against the loop between domains II and III, pS1700 against phosphorylated S1700, pS1928 against phosphorylated S1928, and CNC2 against residues 2122-2138 of α 1 1.2, which are ~40 residues upstream of the very C terminus of α 1 1.2.

    Article Snippet: Concordant with this idea, neither the ACC-003 antibody that we received from Alomone Labs that recognized a 130 kDa band in WT and cKO brains nor our CNC1 antibody recognized the strong 150 kDa band seen with FP1 in brain lysate.

    Techniques:

    Differential recognition of the strong 150 kDa FP1 band in lysate and weak 150 kDa band by FP1, CNC1, and ACC-003 after IP of α 1 1.2 with FP1. Immunoblots with CNC1 ( A , B ), FP1 ( C ), and ACC-003 ( D , E ) of Triton X-100 extracts from WT mice (lysate) and after immunoprecipitation with FP1 from cKO and WT mice. Gels were polymerized from 8% acrylamide. Note that a weak 150 kDa band is detected by CNC1, FP1, and ACC-003 after enrichment of α 1 1.2 by immunoprecipitation with FP1 but the strongly immunoreactive 150 kDa band detected by FP1 in lysate is not detectable by either CNC1 or ACC-003.

    Journal: F1000Research

    Article Title: Proteolytic processing of the L-type Ca2+ channel alpha11.2 subunit in neurons

    doi: 10.12688/f1000research.11808.1

    Figure Lengend Snippet: Differential recognition of the strong 150 kDa FP1 band in lysate and weak 150 kDa band by FP1, CNC1, and ACC-003 after IP of α 1 1.2 with FP1. Immunoblots with CNC1 ( A , B ), FP1 ( C ), and ACC-003 ( D , E ) of Triton X-100 extracts from WT mice (lysate) and after immunoprecipitation with FP1 from cKO and WT mice. Gels were polymerized from 8% acrylamide. Note that a weak 150 kDa band is detected by CNC1, FP1, and ACC-003 after enrichment of α 1 1.2 by immunoprecipitation with FP1 but the strongly immunoreactive 150 kDa band detected by FP1 in lysate is not detectable by either CNC1 or ACC-003.

    Article Snippet: Concordant with this idea, neither the ACC-003 antibody that we received from Alomone Labs that recognized a 130 kDa band in WT and cKO brains nor our CNC1 antibody recognized the strong 150 kDa band seen with FP1 in brain lysate.

    Techniques: Western Blot, Mouse Assay, Immunoprecipitation

    Effect of CAL on I Ca , LTCC expression and the systolic Ca transient. A: Representative L-type Ca currents (I Ca ) recorded at 0 mV from Sham (left) and CAL (right) myocytes. B: Mean (± SEM) I Ca density-voltage relations from Sham (circles, n = 50/15) and CAL (squares, n = 41/15) myocytes. C: Representative Western blots of Sham (S) and CAL lysates probed with antibodies against LTCC (top) or GAPDH (bottom). D: Representative whole-cell Ca transients from Sham (grey line) and CAL (black line) myocytes during field stimulation (left); the right panel shows the early rise on an expanded time scale. E: Mean (± SEM) peak ΔF/F 0 , time to peak (TTP), and rate of rise (ΔF/F 0 .ms − 1 ) of the Ca transient in Sham (open bars, n = 10) and CAL (filled bars, n = 16) myocytes. ⁎⁎⁎ p

    Journal: Journal of Molecular and Cellular Cardiology

    Article Title: Altered distribution of ICa impairs Ca release at the t-tubules of ventricular myocytes from failing hearts

    doi: 10.1016/j.yjmcc.2015.06.012

    Figure Lengend Snippet: Effect of CAL on I Ca , LTCC expression and the systolic Ca transient. A: Representative L-type Ca currents (I Ca ) recorded at 0 mV from Sham (left) and CAL (right) myocytes. B: Mean (± SEM) I Ca density-voltage relations from Sham (circles, n = 50/15) and CAL (squares, n = 41/15) myocytes. C: Representative Western blots of Sham (S) and CAL lysates probed with antibodies against LTCC (top) or GAPDH (bottom). D: Representative whole-cell Ca transients from Sham (grey line) and CAL (black line) myocytes during field stimulation (left); the right panel shows the early rise on an expanded time scale. E: Mean (± SEM) peak ΔF/F 0 , time to peak (TTP), and rate of rise (ΔF/F 0 .ms − 1 ) of the Ca transient in Sham (open bars, n = 10) and CAL (filled bars, n = 16) myocytes. ⁎⁎⁎ p

    Article Snippet: The blot was probed with anti-LTCC antibody (ACC-003; Alomone, Israel) or anti-GAPDH (G9545; Sigma) and protein bands visualized using relevant peroxidase-conjugated secondary antibodies, chemiluminescence, and autoradiography.

    Techniques: Expressing, Western Blot

    CaV1.2 downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets

    doi: 10.1172/JCI124481

    Figure Lengend Snippet: CaV1.2 downregulation mediates impairment of the first phase of insulin secretion in Tardbp knockdown MIN6 cells. ( A ) mRNA expression levels of Cacna1c and Cacna2d1 that encode CaV1.2 and Ca2d1, respectively (control siRNA, Control; Tardbp siRNA, T1 or T2) ( Cacna1c , n = 6; Cacna2d1 , n = 12 each, 1-way ANOVA). ( B ) Immunoblotting of MIN6 cells transfected with control, T1, or T2 (n = 6 each, one-way ANOVA). ( C ) Mock or CaV1.2 plasmid was cotransfected into MIN6 cells with control or T1 siRNA ( n = 4 each, 1-way ANOVA). ( D ) The insulin assay with low and high glucose ( n = 6 each, 1-way ANOVA). ( E ) In situ hybridization of the islets of disease control and ALS subjects with a human CACNA1C mRNA antisense probe. Scale bars: 20 μm. ( F ) Quantitative analysis of in situ hybridization of human CACNA1C , CACNA1D , and CACNA1A ( n = 4 patients each, unpaired t test). Values are mean ± SEM. * P

    Article Snippet: After denaturation, each cell lysate was separated by SDS-PAGE (5%–20% gradient gel) and analyzed by immunoblotting with ECL Plus detection reagents (NEL104001EA, PerkinElmer) using the following primary antibodies: anti–TDP-43 rabbit polyclonal (1:3000; 10782-2-AP, Proteintech), GAPDH mouse monoclonal (1:1000; MBL International Corporation), and anti-CaV1.2 rabbit polyclonal (1:2000; ACC-003, Alomone Labs).

    Techniques: Expressing, Transfection, Plasmid Preparation, In Situ Hybridization

    Loss of TDP-43 reduces insulin secretion by downregulating CaV1.2 calcium channel expression in MIN6 cells.

    Journal: The Journal of Clinical Investigation

    Article Title: TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets

    doi: 10.1172/JCI124481

    Figure Lengend Snippet: Loss of TDP-43 reduces insulin secretion by downregulating CaV1.2 calcium channel expression in MIN6 cells.

    Article Snippet: After denaturation, each cell lysate was separated by SDS-PAGE (5%–20% gradient gel) and analyzed by immunoblotting with ECL Plus detection reagents (NEL104001EA, PerkinElmer) using the following primary antibodies: anti–TDP-43 rabbit polyclonal (1:3000; 10782-2-AP, Proteintech), GAPDH mouse monoclonal (1:1000; MBL International Corporation), and anti-CaV1.2 rabbit polyclonal (1:2000; ACC-003, Alomone Labs).

    Techniques: Expressing

    TDP-43 regulates the transcription of CaV1.2 calcium channels. ( A ) Immunoprecipitation (IP) of lysates from MIN6 cells overexpressing V5-tagged TDP-43 with rabbit IgG or an anti-V5 antibody (IP-V5), analyzed with immunoblotting with the indicated antibody. ( B ) RNA-IP of V5 in cultured MIN6 cells. Bound RNA was analyzed with qRT-PCR using the indicated primers. IP efficiency was calculated relative to input. Representative data from triplicate experiments are shown. ( C ) Schematics of mature mRNA (exon), premature mRNA (intron), and putative promoter are shown over the Cacna1c gene. ( D ) Each mature mRNA (exon) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( E ) mRNA stability was measured in MIN6 cells (control, T1) treated with 10 mg/mL actinomycin D with qRT-PCR using the indicated Cacna1c and Tuba1a primers ( n = 4 each, 2-way ANOVA). The mRNA levels relative to pretreatment were plotted against time after treatment. Residual mRNA levels were compared at 2, 4, 8, and 12 hours after treatment. ( F ) Each premature mRNA (intron) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( G ) Luciferase reporter assay of the –1542/+257 mouse Cacna1c promoter ( n = 10 each, unpaired t test). ( H ) Human CACNA1C promoter luciferase reporter assay using the LightSwitch promoter reporter GoClone collection ( n = 11 each, unpaired t test). Values are mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: TDP-43 regulates early-phase insulin secretion via CaV1.2-mediated exocytosis in islets

    doi: 10.1172/JCI124481

    Figure Lengend Snippet: TDP-43 regulates the transcription of CaV1.2 calcium channels. ( A ) Immunoprecipitation (IP) of lysates from MIN6 cells overexpressing V5-tagged TDP-43 with rabbit IgG or an anti-V5 antibody (IP-V5), analyzed with immunoblotting with the indicated antibody. ( B ) RNA-IP of V5 in cultured MIN6 cells. Bound RNA was analyzed with qRT-PCR using the indicated primers. IP efficiency was calculated relative to input. Representative data from triplicate experiments are shown. ( C ) Schematics of mature mRNA (exon), premature mRNA (intron), and putative promoter are shown over the Cacna1c gene. ( D ) Each mature mRNA (exon) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( E ) mRNA stability was measured in MIN6 cells (control, T1) treated with 10 mg/mL actinomycin D with qRT-PCR using the indicated Cacna1c and Tuba1a primers ( n = 4 each, 2-way ANOVA). The mRNA levels relative to pretreatment were plotted against time after treatment. Residual mRNA levels were compared at 2, 4, 8, and 12 hours after treatment. ( F ) Each premature mRNA (intron) were analyzed with qRT-PCR using the indicated primers ( n = 6 each, unpaired t test). ( G ) Luciferase reporter assay of the –1542/+257 mouse Cacna1c promoter ( n = 10 each, unpaired t test). ( H ) Human CACNA1C promoter luciferase reporter assay using the LightSwitch promoter reporter GoClone collection ( n = 11 each, unpaired t test). Values are mean ± SEM. * P

    Article Snippet: After denaturation, each cell lysate was separated by SDS-PAGE (5%–20% gradient gel) and analyzed by immunoblotting with ECL Plus detection reagents (NEL104001EA, PerkinElmer) using the following primary antibodies: anti–TDP-43 rabbit polyclonal (1:3000; 10782-2-AP, Proteintech), GAPDH mouse monoclonal (1:1000; MBL International Corporation), and anti-CaV1.2 rabbit polyclonal (1:2000; ACC-003, Alomone Labs).

    Techniques: Immunoprecipitation, Cell Culture, Quantitative RT-PCR, Luciferase, Reporter Assay