anti cav1 2 rabbit polyclonal  (Alomone Labs)


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    Name:
    Anti Cav1 2 CACNA1C Antibody
    Description:
    Anti CaV1 2 CACNA1C Antibody ACC 003 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunoprecipitation immunohistochemistry immunocytochemistry and indirect flow cytometry applications It has been designed to recognize CaV1 2 from mouse rat and human samples
    Catalog Number:
    ACC-003
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Indirect Flow Cytometry, Immunohistochemistry, Immunoprecipitation, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
    Buy from Supplier


    Structured Review

    Alomone Labs anti cav1 2 rabbit polyclonal
    Anti Cav1 2 CACNA1C Antibody
    Anti CaV1 2 CACNA1C Antibody ACC 003 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunoprecipitation immunohistochemistry immunocytochemistry and indirect flow cytometry applications It has been designed to recognize CaV1 2 from mouse rat and human samples
    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, 2021-10
    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

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

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

    Journal: Journal of Neurochemistry

    doi: 10.1111/j.1471-4159.2010.07089.x

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

    Techniques Used: Isolation, Mouse Assay, Labeling

    Related Articles

    Immunofluorescence:

    Article Title: Attenuated β-adrenergic response in calcium/calmodulin-dependent protein kinase IV-knockout mice
    Article Snippet: .. Whole-cell immunofluorescence labeling of isolated cardiac myocytes was performed using an anti-CaV1.2 antibody at a dilution of 1:100 (Alomone Labs, Jerusalem, Israel) as the primary antibody; visualization was performed using fluorescein-conjugated polyclonal goat anti-guinea pig IgG at a dilution of 1:200. ..

    Labeling:

    Article Title: Attenuated β-adrenergic response in calcium/calmodulin-dependent protein kinase IV-knockout mice
    Article Snippet: .. Whole-cell immunofluorescence labeling of isolated cardiac myocytes was performed using an anti-CaV1.2 antibody at a dilution of 1:100 (Alomone Labs, Jerusalem, Israel) as the primary antibody; visualization was performed using fluorescein-conjugated polyclonal goat anti-guinea pig IgG at a dilution of 1:200. ..

    Isolation:

    Article Title: Attenuated β-adrenergic response in calcium/calmodulin-dependent protein kinase IV-knockout mice
    Article Snippet: .. Whole-cell immunofluorescence labeling of isolated cardiac myocytes was performed using an anti-CaV1.2 antibody at a dilution of 1:100 (Alomone Labs, Jerusalem, Israel) as the primary antibody; visualization was performed using fluorescein-conjugated polyclonal goat anti-guinea pig IgG at a dilution of 1:200. ..

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  • 95
    Alomone Labs anti cav1 2
    Expressions of calcium handling proteins. The confocal image showed fluorescein isothiocyanate (green)-labeled <t>Cav1.2,</t> SERCA2a, and NCX1 expressed in isolated rat cardiac ventricular myocytes in SO (left panel), HF (middle panel), and NRG group (right panel). Nuclei were stained with DAPI (blue). The scale bar represents 20 μm (n = 5 for each set of staining).
    Anti Cav1 2, 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/anti cav1 2/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 - by Bioz Stars, 2021-10
    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, 2021-10
    86/100 stars
      Buy from Supplier

    Image Search Results


    Expressions of calcium handling proteins. The confocal image showed fluorescein isothiocyanate (green)-labeled Cav1.2, SERCA2a, and NCX1 expressed in isolated rat cardiac ventricular myocytes in SO (left panel), HF (middle panel), and NRG group (right panel). Nuclei were stained with DAPI (blue). The scale bar represents 20 μm (n = 5 for each set of staining).

    Journal: Frontiers in Pharmacology

    Article Title: Neuregulin-1β Partially Improves Cardiac Function in Volume-Overload Heart Failure Through Regulation of Abnormal Calcium Handling

    doi: 10.3389/fphar.2019.00616

    Figure Lengend Snippet: Expressions of calcium handling proteins. The confocal image showed fluorescein isothiocyanate (green)-labeled Cav1.2, SERCA2a, and NCX1 expressed in isolated rat cardiac ventricular myocytes in SO (left panel), HF (middle panel), and NRG group (right panel). Nuclei were stained with DAPI (blue). The scale bar represents 20 μm (n = 5 for each set of staining).

    Article Snippet: Anti-Cav1.2, anti-SERCA2a, and anti-NCX1 antibody were purchased from Alomone (Jerusalem, ACC-003, Israel), Millipore (MA, AB3516P, USA), and Santa Cruz Biotechnology (CA, SC-8094, USA), and secondary antibodies, anti-rabbit488 conjugated antibody was purchased from Santa Cruz Biotechnology (CA, sc-362262, USA).

    Techniques: Labeling, Isolation, Staining

    Quantitative analysis of calcium handling proteins expressions. (A) Representative expression of Cav1.2, SERCA2a, and NCX1, β-actin as loading controls. (B) Comparison of Cav1.2 protein expression (n = 3 rats). (C) Comparison of SERCA2a protein expression (n = 4 rats). (D) Comparison of NCX1 protein expression (n = 4 rats). * P

    Journal: Frontiers in Pharmacology

    Article Title: Neuregulin-1β Partially Improves Cardiac Function in Volume-Overload Heart Failure Through Regulation of Abnormal Calcium Handling

    doi: 10.3389/fphar.2019.00616

    Figure Lengend Snippet: Quantitative analysis of calcium handling proteins expressions. (A) Representative expression of Cav1.2, SERCA2a, and NCX1, β-actin as loading controls. (B) Comparison of Cav1.2 protein expression (n = 3 rats). (C) Comparison of SERCA2a protein expression (n = 4 rats). (D) Comparison of NCX1 protein expression (n = 4 rats). * P

    Article Snippet: Anti-Cav1.2, anti-SERCA2a, and anti-NCX1 antibody were purchased from Alomone (Jerusalem, ACC-003, Israel), Millipore (MA, AB3516P, USA), and Santa Cruz Biotechnology (CA, SC-8094, USA), and secondary antibodies, anti-rabbit488 conjugated antibody was purchased from Santa Cruz Biotechnology (CA, sc-362262, USA).

    Techniques: Expressing

    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