rabbit anti β3  (Alomone Labs)


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    Alomone Labs rabbit anti β3
    Interaction between pore-forming Ca V 1.3 and auxiliary β-subunits of voltage-dependent Ca 2+ channels. 1A: Left panel : CHO cells were transfected with Ca V 1.3-GFP fusion construct and <t>β3-subunits,</t> precipitated with antibodies against β3-subunits and blotted for Ca V 1.3 protein. Proteins precipitated using antibodies against GFP were positively stained with antibodies against Ca V 1.3 indicating identification of Ca V 1.3 subunits. Right two panels : CHO cells transfected with Ca V 1.3 and β3-subunits; Ca V 1.3 was immunoprecipitated and blotted for β3-subunits. Control experiment: precipitation using anti-β3-antibody and blot stained against β3-subunit. (L = lysate, 10% of total protein; IP = immunoprecipitation) 1B : ARPE-19 cells transfected with β3-subunit (green) and Ca V 1.3 (red). The merged picture shows co-localization of β3-subunits and Ca V 1.3. On the right: fluorescence profile showing subcellular protein distribution. 1C : To quantify plasma membrane localization, pixel analysis was performed for edge detection to calculate surface expression (data are mean ± SEM; n = 3). Scale bar represents 10 µm.
    Rabbit Anti β3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti β3/product/Alomone Labs
    Average 94 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    rabbit anti β3 - by Bioz Stars, 2022-08
    94/100 stars

    Images

    1) Product Images from "Interaction of Bestrophin-1 and Ca2+ Channel ?-Subunits: Identification of New Binding Domains on the Bestrophin-1 C-Terminus"

    Article Title: Interaction of Bestrophin-1 and Ca2+ Channel ?-Subunits: Identification of New Binding Domains on the Bestrophin-1 C-Terminus

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0019364

    Interaction between pore-forming Ca V 1.3 and auxiliary β-subunits of voltage-dependent Ca 2+ channels. 1A: Left panel : CHO cells were transfected with Ca V 1.3-GFP fusion construct and β3-subunits, precipitated with antibodies against β3-subunits and blotted for Ca V 1.3 protein. Proteins precipitated using antibodies against GFP were positively stained with antibodies against Ca V 1.3 indicating identification of Ca V 1.3 subunits. Right two panels : CHO cells transfected with Ca V 1.3 and β3-subunits; Ca V 1.3 was immunoprecipitated and blotted for β3-subunits. Control experiment: precipitation using anti-β3-antibody and blot stained against β3-subunit. (L = lysate, 10% of total protein; IP = immunoprecipitation) 1B : ARPE-19 cells transfected with β3-subunit (green) and Ca V 1.3 (red). The merged picture shows co-localization of β3-subunits and Ca V 1.3. On the right: fluorescence profile showing subcellular protein distribution. 1C : To quantify plasma membrane localization, pixel analysis was performed for edge detection to calculate surface expression (data are mean ± SEM; n = 3). Scale bar represents 10 µm.
    Figure Legend Snippet: Interaction between pore-forming Ca V 1.3 and auxiliary β-subunits of voltage-dependent Ca 2+ channels. 1A: Left panel : CHO cells were transfected with Ca V 1.3-GFP fusion construct and β3-subunits, precipitated with antibodies against β3-subunits and blotted for Ca V 1.3 protein. Proteins precipitated using antibodies against GFP were positively stained with antibodies against Ca V 1.3 indicating identification of Ca V 1.3 subunits. Right two panels : CHO cells transfected with Ca V 1.3 and β3-subunits; Ca V 1.3 was immunoprecipitated and blotted for β3-subunits. Control experiment: precipitation using anti-β3-antibody and blot stained against β3-subunit. (L = lysate, 10% of total protein; IP = immunoprecipitation) 1B : ARPE-19 cells transfected with β3-subunit (green) and Ca V 1.3 (red). The merged picture shows co-localization of β3-subunits and Ca V 1.3. On the right: fluorescence profile showing subcellular protein distribution. 1C : To quantify plasma membrane localization, pixel analysis was performed for edge detection to calculate surface expression (data are mean ± SEM; n = 3). Scale bar represents 10 µm.

    Techniques Used: Transfection, Construct, Staining, Immunoprecipitation, Fluorescence, Expressing

    Subcellular localization of heterologously expressed Ca V 1.3, β-subunits of voltage-dependent Ca 2+ channels and bestrophin-1: ARPE-19 cells were transfected with: β3-subunits and bestrophin-1, Ca V 1.3, β3 or β4-subunits and bestrophin-1, and human P2Y 2 -His 6 receptor. 2A : Cells transfected with β3-subunit (green) and bestrophin-1 (red). Yellow colour in the merged picture indicates interaction of both proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2B : Cells transfected with β3-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2C : Cells transfected with β4-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2D : Human P2Y 2 -His 6 receptor which shows plasma membrane localization as a control. Note: cells which express Ca V 1.3 and β-subunits always appear in a more spherical shape and do not remain flat due to the expression of the large L-type channel subunits. The smaller P2Y 2 -receptor did not change the cell shape. 2E : Relative surface expression quantified by edge detection analysis (data are mean ± SEM; n = 3). (* = p
    Figure Legend Snippet: Subcellular localization of heterologously expressed Ca V 1.3, β-subunits of voltage-dependent Ca 2+ channels and bestrophin-1: ARPE-19 cells were transfected with: β3-subunits and bestrophin-1, Ca V 1.3, β3 or β4-subunits and bestrophin-1, and human P2Y 2 -His 6 receptor. 2A : Cells transfected with β3-subunit (green) and bestrophin-1 (red). Yellow colour in the merged picture indicates interaction of both proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2B : Cells transfected with β3-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2C : Cells transfected with β4-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2D : Human P2Y 2 -His 6 receptor which shows plasma membrane localization as a control. Note: cells which express Ca V 1.3 and β-subunits always appear in a more spherical shape and do not remain flat due to the expression of the large L-type channel subunits. The smaller P2Y 2 -receptor did not change the cell shape. 2E : Relative surface expression quantified by edge detection analysis (data are mean ± SEM; n = 3). (* = p

    Techniques Used: Transfection, Fluorescence, Expressing

    Detection of interaction sites between β-subunits and bestrophin-1. 3A : Bestrophin-1 construct used in this study and alignment of amino acid sequences of the C-terminus of the bestrophin-1 from different species (Boxes: transmembrane domains). Two among vertebrate species highly conserved clusters of proline-rich motifs (PxxP) could be detected. In the ΔCTPxxP mutant form, PxxP motifs between amino acid 468 to 486 were removed with unchanged recognition sites for the anti bestrophin-1 antibody. 3B : HEK-293 cells were transfected with β3-subunits together with bestrophin-1 or ΔCTPxxP constructs. Proteins were precipitated using anti-bestrophin-1 antibody and blots were visualized for anti-β3-subunit to show co-immunoprecipitation. 3C : HEK-293 cells were transfected with His-tagged β4-subunits together with bestrophin-1 wild type or ΔCTPxxP constructs. Proteins were precipitated using anti-His antibody and the blots were visualized with anti-bestrophin-1 antibody to show co-immunoprecipitation. 3D : Relative co-immunoprecipitation of β3-subunits with either wild-type or ΔCTPxxP bestrophin-1: efficiency was measured by densitometry (n = 5). 3E : Relative co-immunoprecipitation of β4-subunits with either wild-type or ΔCTPxxP bestrophin-1 (depicted n = 3). (L = lysate; IP = immunoprecipitation; NB = not bound). The following species abbreviations were used: Hs, Homo sapiens, Mm, Macaca mulatta, Bt, Bos taurus, Ms, Mus musculus, Xt, Xenopus tropicalis, Fr, Fugu rubripes, and Ci, Ciona intestinalis.
    Figure Legend Snippet: Detection of interaction sites between β-subunits and bestrophin-1. 3A : Bestrophin-1 construct used in this study and alignment of amino acid sequences of the C-terminus of the bestrophin-1 from different species (Boxes: transmembrane domains). Two among vertebrate species highly conserved clusters of proline-rich motifs (PxxP) could be detected. In the ΔCTPxxP mutant form, PxxP motifs between amino acid 468 to 486 were removed with unchanged recognition sites for the anti bestrophin-1 antibody. 3B : HEK-293 cells were transfected with β3-subunits together with bestrophin-1 or ΔCTPxxP constructs. Proteins were precipitated using anti-bestrophin-1 antibody and blots were visualized for anti-β3-subunit to show co-immunoprecipitation. 3C : HEK-293 cells were transfected with His-tagged β4-subunits together with bestrophin-1 wild type or ΔCTPxxP constructs. Proteins were precipitated using anti-His antibody and the blots were visualized with anti-bestrophin-1 antibody to show co-immunoprecipitation. 3D : Relative co-immunoprecipitation of β3-subunits with either wild-type or ΔCTPxxP bestrophin-1: efficiency was measured by densitometry (n = 5). 3E : Relative co-immunoprecipitation of β4-subunits with either wild-type or ΔCTPxxP bestrophin-1 (depicted n = 3). (L = lysate; IP = immunoprecipitation; NB = not bound). The following species abbreviations were used: Hs, Homo sapiens, Mm, Macaca mulatta, Bt, Bos taurus, Ms, Mus musculus, Xt, Xenopus tropicalis, Fr, Fugu rubripes, and Ci, Ciona intestinalis.

    Techniques Used: Construct, Mutagenesis, Transfection, Immunoprecipitation, Mass Spectrometry

    2) Product Images from "CACNB4 overexpression decreases dendritic spine density in sex-specific manner"

    Article Title: CACNB4 overexpression decreases dendritic spine density in sex-specific manner

    Journal: bioRxiv

    doi: 10.1101/2022.02.02.478824

    β subunit protein levels. A) (Left) Mean optical density (a.u.) of β4 was significantly lower in P84, relative to P28 male and female mice (F=9.635, DF=3, p=0.013), with no significant sex or age by sex interactions. Error bars = SEM. (Right) Representative blot showing β4 bands at ~51-55kD. B) (Left) Mean optical density (a.u.) of β1 was significantly lower in P84, relative to P28 male and female mice (F=21.499, DF=3, p=0.001). There was also a main effect of sex (F=6.944, DF=3, p=0.027) and a significant age by sex interaction (F=6.835, DF=3, p=0 .028). Main effect of sex was significant at P28 (F=12.138, DF=1, p=0.040) but not P84 (F=0.016, DF=1, p=0.906). Error bars = SEM. (Right) Representative blot showing β1 bands at ~50-80kD. C) (Left) Neither age nor sex significantly impacted mean optical density (a.u.) of β3 and there was not a significant sex by age interaction. Error bars = SEM. (Right) Representative blot showing β3 bands at ~55kD (molecular weight of the predominant β3 isoform).
    Figure Legend Snippet: β subunit protein levels. A) (Left) Mean optical density (a.u.) of β4 was significantly lower in P84, relative to P28 male and female mice (F=9.635, DF=3, p=0.013), with no significant sex or age by sex interactions. Error bars = SEM. (Right) Representative blot showing β4 bands at ~51-55kD. B) (Left) Mean optical density (a.u.) of β1 was significantly lower in P84, relative to P28 male and female mice (F=21.499, DF=3, p=0.001). There was also a main effect of sex (F=6.944, DF=3, p=0.027) and a significant age by sex interaction (F=6.835, DF=3, p=0 .028). Main effect of sex was significant at P28 (F=12.138, DF=1, p=0.040) but not P84 (F=0.016, DF=1, p=0.906). Error bars = SEM. (Right) Representative blot showing β1 bands at ~50-80kD. C) (Left) Neither age nor sex significantly impacted mean optical density (a.u.) of β3 and there was not a significant sex by age interaction. Error bars = SEM. (Right) Representative blot showing β3 bands at ~55kD (molecular weight of the predominant β3 isoform).

    Techniques Used: Mouse Assay, Molecular Weight

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    Alomone Labs rabbit anti β3
    Interaction between pore-forming Ca V 1.3 and auxiliary β-subunits of voltage-dependent Ca 2+ channels. 1A: Left panel : CHO cells were transfected with Ca V 1.3-GFP fusion construct and <t>β3-subunits,</t> precipitated with antibodies against β3-subunits and blotted for Ca V 1.3 protein. Proteins precipitated using antibodies against GFP were positively stained with antibodies against Ca V 1.3 indicating identification of Ca V 1.3 subunits. Right two panels : CHO cells transfected with Ca V 1.3 and β3-subunits; Ca V 1.3 was immunoprecipitated and blotted for β3-subunits. Control experiment: precipitation using anti-β3-antibody and blot stained against β3-subunit. (L = lysate, 10% of total protein; IP = immunoprecipitation) 1B : ARPE-19 cells transfected with β3-subunit (green) and Ca V 1.3 (red). The merged picture shows co-localization of β3-subunits and Ca V 1.3. On the right: fluorescence profile showing subcellular protein distribution. 1C : To quantify plasma membrane localization, pixel analysis was performed for edge detection to calculate surface expression (data are mean ± SEM; n = 3). Scale bar represents 10 µm.
    Rabbit Anti β3, 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
    https://www.bioz.com/result/rabbit anti β3/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti β3 - by Bioz Stars, 2022-08
    94/100 stars
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    Interaction between pore-forming Ca V 1.3 and auxiliary β-subunits of voltage-dependent Ca 2+ channels. 1A: Left panel : CHO cells were transfected with Ca V 1.3-GFP fusion construct and β3-subunits, precipitated with antibodies against β3-subunits and blotted for Ca V 1.3 protein. Proteins precipitated using antibodies against GFP were positively stained with antibodies against Ca V 1.3 indicating identification of Ca V 1.3 subunits. Right two panels : CHO cells transfected with Ca V 1.3 and β3-subunits; Ca V 1.3 was immunoprecipitated and blotted for β3-subunits. Control experiment: precipitation using anti-β3-antibody and blot stained against β3-subunit. (L = lysate, 10% of total protein; IP = immunoprecipitation) 1B : ARPE-19 cells transfected with β3-subunit (green) and Ca V 1.3 (red). The merged picture shows co-localization of β3-subunits and Ca V 1.3. On the right: fluorescence profile showing subcellular protein distribution. 1C : To quantify plasma membrane localization, pixel analysis was performed for edge detection to calculate surface expression (data are mean ± SEM; n = 3). Scale bar represents 10 µm.

    Journal: PLoS ONE

    Article Title: Interaction of Bestrophin-1 and Ca2+ Channel ?-Subunits: Identification of New Binding Domains on the Bestrophin-1 C-Terminus

    doi: 10.1371/journal.pone.0019364

    Figure Lengend Snippet: Interaction between pore-forming Ca V 1.3 and auxiliary β-subunits of voltage-dependent Ca 2+ channels. 1A: Left panel : CHO cells were transfected with Ca V 1.3-GFP fusion construct and β3-subunits, precipitated with antibodies against β3-subunits and blotted for Ca V 1.3 protein. Proteins precipitated using antibodies against GFP were positively stained with antibodies against Ca V 1.3 indicating identification of Ca V 1.3 subunits. Right two panels : CHO cells transfected with Ca V 1.3 and β3-subunits; Ca V 1.3 was immunoprecipitated and blotted for β3-subunits. Control experiment: precipitation using anti-β3-antibody and blot stained against β3-subunit. (L = lysate, 10% of total protein; IP = immunoprecipitation) 1B : ARPE-19 cells transfected with β3-subunit (green) and Ca V 1.3 (red). The merged picture shows co-localization of β3-subunits and Ca V 1.3. On the right: fluorescence profile showing subcellular protein distribution. 1C : To quantify plasma membrane localization, pixel analysis was performed for edge detection to calculate surface expression (data are mean ± SEM; n = 3). Scale bar represents 10 µm.

    Article Snippet: Antibodies Proteins were detected using the following antibodies: mouse monoclonal anti-human-bestrophin ab2182, mouse monoclonal anti His6 ab18184, rabbit polyclonal anti His6ab9108 (Abcam plc, Cambridge, UK), rabbit anti-β3, rabbit Cav1.3 (Alomone Labs), goat anti Cav1.3 (Santa Cruz), mouse monoclonal anti beta-actin (clone JLA20, Hybridoma Bank Iowa), and mouse monoclonal anti-GFP (Roche).

    Techniques: Transfection, Construct, Staining, Immunoprecipitation, Fluorescence, Expressing

    Subcellular localization of heterologously expressed Ca V 1.3, β-subunits of voltage-dependent Ca 2+ channels and bestrophin-1: ARPE-19 cells were transfected with: β3-subunits and bestrophin-1, Ca V 1.3, β3 or β4-subunits and bestrophin-1, and human P2Y 2 -His 6 receptor. 2A : Cells transfected with β3-subunit (green) and bestrophin-1 (red). Yellow colour in the merged picture indicates interaction of both proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2B : Cells transfected with β3-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2C : Cells transfected with β4-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2D : Human P2Y 2 -His 6 receptor which shows plasma membrane localization as a control. Note: cells which express Ca V 1.3 and β-subunits always appear in a more spherical shape and do not remain flat due to the expression of the large L-type channel subunits. The smaller P2Y 2 -receptor did not change the cell shape. 2E : Relative surface expression quantified by edge detection analysis (data are mean ± SEM; n = 3). (* = p

    Journal: PLoS ONE

    Article Title: Interaction of Bestrophin-1 and Ca2+ Channel ?-Subunits: Identification of New Binding Domains on the Bestrophin-1 C-Terminus

    doi: 10.1371/journal.pone.0019364

    Figure Lengend Snippet: Subcellular localization of heterologously expressed Ca V 1.3, β-subunits of voltage-dependent Ca 2+ channels and bestrophin-1: ARPE-19 cells were transfected with: β3-subunits and bestrophin-1, Ca V 1.3, β3 or β4-subunits and bestrophin-1, and human P2Y 2 -His 6 receptor. 2A : Cells transfected with β3-subunit (green) and bestrophin-1 (red). Yellow colour in the merged picture indicates interaction of both proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2B : Cells transfected with β3-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2C : Cells transfected with β4-subunit (green), bestrophin-1 (red), and Ca v 1.3 subunit (blue). White colour in the merged picture suggests co-localization of all three proteins. On the right: fluorescence profiles showing subcellular protein distribution. 2D : Human P2Y 2 -His 6 receptor which shows plasma membrane localization as a control. Note: cells which express Ca V 1.3 and β-subunits always appear in a more spherical shape and do not remain flat due to the expression of the large L-type channel subunits. The smaller P2Y 2 -receptor did not change the cell shape. 2E : Relative surface expression quantified by edge detection analysis (data are mean ± SEM; n = 3). (* = p

    Article Snippet: Antibodies Proteins were detected using the following antibodies: mouse monoclonal anti-human-bestrophin ab2182, mouse monoclonal anti His6 ab18184, rabbit polyclonal anti His6ab9108 (Abcam plc, Cambridge, UK), rabbit anti-β3, rabbit Cav1.3 (Alomone Labs), goat anti Cav1.3 (Santa Cruz), mouse monoclonal anti beta-actin (clone JLA20, Hybridoma Bank Iowa), and mouse monoclonal anti-GFP (Roche).

    Techniques: Transfection, Fluorescence, Expressing

    Detection of interaction sites between β-subunits and bestrophin-1. 3A : Bestrophin-1 construct used in this study and alignment of amino acid sequences of the C-terminus of the bestrophin-1 from different species (Boxes: transmembrane domains). Two among vertebrate species highly conserved clusters of proline-rich motifs (PxxP) could be detected. In the ΔCTPxxP mutant form, PxxP motifs between amino acid 468 to 486 were removed with unchanged recognition sites for the anti bestrophin-1 antibody. 3B : HEK-293 cells were transfected with β3-subunits together with bestrophin-1 or ΔCTPxxP constructs. Proteins were precipitated using anti-bestrophin-1 antibody and blots were visualized for anti-β3-subunit to show co-immunoprecipitation. 3C : HEK-293 cells were transfected with His-tagged β4-subunits together with bestrophin-1 wild type or ΔCTPxxP constructs. Proteins were precipitated using anti-His antibody and the blots were visualized with anti-bestrophin-1 antibody to show co-immunoprecipitation. 3D : Relative co-immunoprecipitation of β3-subunits with either wild-type or ΔCTPxxP bestrophin-1: efficiency was measured by densitometry (n = 5). 3E : Relative co-immunoprecipitation of β4-subunits with either wild-type or ΔCTPxxP bestrophin-1 (depicted n = 3). (L = lysate; IP = immunoprecipitation; NB = not bound). The following species abbreviations were used: Hs, Homo sapiens, Mm, Macaca mulatta, Bt, Bos taurus, Ms, Mus musculus, Xt, Xenopus tropicalis, Fr, Fugu rubripes, and Ci, Ciona intestinalis.

    Journal: PLoS ONE

    Article Title: Interaction of Bestrophin-1 and Ca2+ Channel ?-Subunits: Identification of New Binding Domains on the Bestrophin-1 C-Terminus

    doi: 10.1371/journal.pone.0019364

    Figure Lengend Snippet: Detection of interaction sites between β-subunits and bestrophin-1. 3A : Bestrophin-1 construct used in this study and alignment of amino acid sequences of the C-terminus of the bestrophin-1 from different species (Boxes: transmembrane domains). Two among vertebrate species highly conserved clusters of proline-rich motifs (PxxP) could be detected. In the ΔCTPxxP mutant form, PxxP motifs between amino acid 468 to 486 were removed with unchanged recognition sites for the anti bestrophin-1 antibody. 3B : HEK-293 cells were transfected with β3-subunits together with bestrophin-1 or ΔCTPxxP constructs. Proteins were precipitated using anti-bestrophin-1 antibody and blots were visualized for anti-β3-subunit to show co-immunoprecipitation. 3C : HEK-293 cells were transfected with His-tagged β4-subunits together with bestrophin-1 wild type or ΔCTPxxP constructs. Proteins were precipitated using anti-His antibody and the blots were visualized with anti-bestrophin-1 antibody to show co-immunoprecipitation. 3D : Relative co-immunoprecipitation of β3-subunits with either wild-type or ΔCTPxxP bestrophin-1: efficiency was measured by densitometry (n = 5). 3E : Relative co-immunoprecipitation of β4-subunits with either wild-type or ΔCTPxxP bestrophin-1 (depicted n = 3). (L = lysate; IP = immunoprecipitation; NB = not bound). The following species abbreviations were used: Hs, Homo sapiens, Mm, Macaca mulatta, Bt, Bos taurus, Ms, Mus musculus, Xt, Xenopus tropicalis, Fr, Fugu rubripes, and Ci, Ciona intestinalis.

    Article Snippet: Antibodies Proteins were detected using the following antibodies: mouse monoclonal anti-human-bestrophin ab2182, mouse monoclonal anti His6 ab18184, rabbit polyclonal anti His6ab9108 (Abcam plc, Cambridge, UK), rabbit anti-β3, rabbit Cav1.3 (Alomone Labs), goat anti Cav1.3 (Santa Cruz), mouse monoclonal anti beta-actin (clone JLA20, Hybridoma Bank Iowa), and mouse monoclonal anti-GFP (Roche).

    Techniques: Construct, Mutagenesis, Transfection, Immunoprecipitation, Mass Spectrometry

    β subunit protein levels. A) (Left) Mean optical density (a.u.) of β4 was significantly lower in P84, relative to P28 male and female mice (F=9.635, DF=3, p=0.013), with no significant sex or age by sex interactions. Error bars = SEM. (Right) Representative blot showing β4 bands at ~51-55kD. B) (Left) Mean optical density (a.u.) of β1 was significantly lower in P84, relative to P28 male and female mice (F=21.499, DF=3, p=0.001). There was also a main effect of sex (F=6.944, DF=3, p=0.027) and a significant age by sex interaction (F=6.835, DF=3, p=0 .028). Main effect of sex was significant at P28 (F=12.138, DF=1, p=0.040) but not P84 (F=0.016, DF=1, p=0.906). Error bars = SEM. (Right) Representative blot showing β1 bands at ~50-80kD. C) (Left) Neither age nor sex significantly impacted mean optical density (a.u.) of β3 and there was not a significant sex by age interaction. Error bars = SEM. (Right) Representative blot showing β3 bands at ~55kD (molecular weight of the predominant β3 isoform).

    Journal: bioRxiv

    Article Title: CACNB4 overexpression decreases dendritic spine density in sex-specific manner

    doi: 10.1101/2022.02.02.478824

    Figure Lengend Snippet: β subunit protein levels. A) (Left) Mean optical density (a.u.) of β4 was significantly lower in P84, relative to P28 male and female mice (F=9.635, DF=3, p=0.013), with no significant sex or age by sex interactions. Error bars = SEM. (Right) Representative blot showing β4 bands at ~51-55kD. B) (Left) Mean optical density (a.u.) of β1 was significantly lower in P84, relative to P28 male and female mice (F=21.499, DF=3, p=0.001). There was also a main effect of sex (F=6.944, DF=3, p=0.027) and a significant age by sex interaction (F=6.835, DF=3, p=0 .028). Main effect of sex was significant at P28 (F=12.138, DF=1, p=0.040) but not P84 (F=0.016, DF=1, p=0.906). Error bars = SEM. (Right) Representative blot showing β1 bands at ~50-80kD. C) (Left) Neither age nor sex significantly impacted mean optical density (a.u.) of β3 and there was not a significant sex by age interaction. Error bars = SEM. (Right) Representative blot showing β3 bands at ~55kD (molecular weight of the predominant β3 isoform).

    Article Snippet: The membrane was incubated overnight in Pierce SuperBlock Blocking Buffer and validated primary antibodies: either mouse anti-CaV β1 (Neuromab #73-052 1:100), rabbit anti-CaV β3 (Alomone Labs #AAC-008, 1:800) or mouse anti-CaVβ4 (Neuromab #75-054, 1:1000) and rabbit anti-β-tubulin (Abcam #ab6046, 1:600,000).

    Techniques: Mouse Assay, Molecular Weight