alpha-sarcoglycan Search Results


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    Thermo Fisher gene exp sgcd hs01087180 m1
    Expression of <t>S151A-SGCD</t> precedes cardiac function changes in adult Drosophila expressing S151A-SGCD. (A).Temperature shift from 18°C to 26°C results in deterioration in cardiac function and induction of S151A-SGCD expression. After a temperature shift to 26°C, the induction of transgene expression precedes the development of dilated cardiomyopathy by 48 hours as demonstrated by QRT-PCR measurements of S151A-SGCD mRNA levels compared to cardiac function by serial OCT. A second temperature shift back to 18°C represses S151A-SGCD mRNA expression and results in restoration of cardiac function by 48 hours. We performed three independent experiments using different batches of flies each time. The summary data for QRT-PCR at the indicated times and temperatures are expressed as the mean +/− SE of three independent experiments, each performed in triplicate. * p
    Gene Exp Sgcd Hs01087180 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Leica Biosystems alpha sarcoglycan
    Expression of <t>S151A-SGCD</t> precedes cardiac function changes in adult Drosophila expressing S151A-SGCD. (A).Temperature shift from 18°C to 26°C results in deterioration in cardiac function and induction of S151A-SGCD expression. After a temperature shift to 26°C, the induction of transgene expression precedes the development of dilated cardiomyopathy by 48 hours as demonstrated by QRT-PCR measurements of S151A-SGCD mRNA levels compared to cardiac function by serial OCT. A second temperature shift back to 18°C represses S151A-SGCD mRNA expression and results in restoration of cardiac function by 48 hours. We performed three independent experiments using different batches of flies each time. The summary data for QRT-PCR at the indicated times and temperatures are expressed as the mean +/− SE of three independent experiments, each performed in triplicate. * p
    Alpha Sarcoglycan, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    MONOSAN biotinylated anti alpha sarcoglycan
    Isolation of <t>alpha-sarcoglycan</t> + EVs from plasma using immunoaffinity capturing. Anti-alpha-sarcoglycan antibodies were conjugated to magnetic beads to isolate muscle EVs from plasma. Western blot analysis confirmed the presence of the exosomal marker Tsg101 in isolated EVs. Ponceau S Staining has been used as loading control (a). MicroRNA quantifications showed an increase in the miR-206/miR-16 ratio in the SGCA + sub-population of EVs (SGCA-Beads) compared to total (total plasma EVs) or uncaptured EVs (Supernatant). MiR-206 expression levels were normalized versus the endogenous reference miR-16 and expressed as -ΔCq (where ΔCq = Cq miR-206 -Cq miR-16 ) (b). Moreover, the quantification of miR-16 ratio in the SGCA + sub-population of EVs compared to total or uncaptured EVs shows that SGCA-conjugated beads retained about 2–5% of the total amount of EVs, miR-16 expression levels were normalized versus the spike-in reference cel-miR-39 and expressed as -ΔCq (where ΔCq = Cq miR-16 -Cq cel-miR-39 ) (c). Asterisks denote significant changes ( p
    Biotinylated Anti Alpha Sarcoglycan, supplied by MONOSAN, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novocastra alpha sarcoglycan
    Cross-sectional profiles of regenerating fibres. Three serial frozen cross sections of a biopsy obtained from regenerating vastus lateralis skeletal muscle 7 days after injury induced by electrical stimulation-elicited eccentric contractions (right). Three serial cross sections from the control (uninjured) leg of the same individual are shown (left) for comparison. The sections have been stained with <t>alpha-sarcoglycan,</t> beta-dystroglycan or dystrophin to label the sarcolemma, along with a basement membrane protein (laminin or collagen IV) and a myogenic marker (desmin or neonatal/embryonic myosin; MHCn/e). Each column of images contains single channel and combined images for each staining. In the injured muscle, dystrophin staining is completely absent in several fibres, while the basement membrane (laminin) is preserved. MHCn/e staining is evident in some small dystrophin-negative fibres. A similar pattern is evident from the alpha-sarcoglycan and beta-dystroglycan staining, with negative fibres, together with desmin+ cells, contained within a preserved basement membrane (collagen IV). Asterisk indicates some of the necrotic fibres. Note the different profiles of the injured fibres, such as varying fibre size, and infiltrating cells either confined to the fibre periphery or dispersed throughout the fibre. Scale bar, 100 μm
    Alpha Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    sgα  (Abcam)
    77
    Abcam sgα
    Ameliorated NMJ fragmentation and denervation in aged mice by <t>SGα</t> overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.
    Sgα, supplied by Abcam, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Journal of Biological Chemistry sarcoglycan adhalin
    Ameliorated NMJ fragmentation and denervation in aged mice by <t>SGα</t> overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.
    Sarcoglycan Adhalin, supplied by Journal of Biological Chemistry, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novocastra α sarcoglycan adhalin
    Ameliorated NMJ fragmentation and denervation in aged mice by <t>SGα</t> overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.
    α Sarcoglycan Adhalin, supplied by Novocastra, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novocastra mouse monoclonal anti adhalin antibody
    Ameliorated NMJ fragmentation and denervation in aged mice by <t>SGα</t> overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.
    Mouse Monoclonal Anti Adhalin Antibody, supplied by Novocastra, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Novocastra sarcoglycans
    Ameliorated NMJ fragmentation and denervation in aged mice by <t>SGα</t> overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.
    Sarcoglycans, supplied by Novocastra, used in various techniques. Bioz Stars score: 87/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals beta sarcoglycan antibody
    Ameliorated NMJ fragmentation and denervation in aged mice by <t>SGα</t> overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.
    Beta Sarcoglycan Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 82/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leica Biosystems mouse monoclonal beta sarcoglycan antibodies
    Morphological and molecular evidences of myocardial hypertrophy progression. (A) Immunohistochemical staining of <t>beta-</t> <t>sarcoglycan</t> allowing to detect outer membrane showing clear increase of cells size in transverse orientation by 10-weeks group compared to intact. (B) Increase in cell diameter (Dmin) illustrating progressive cardiomyocytes enlargement, and significant increase after 8 and 10 weeks of aortic banding performing. Nppa expression was upregulated in LV (C) and IVS (D) after 1, 2, 8, and 10 weeks of model duration compared to intact or sham-control groups. (E) Positive linear correlation was found for Nppa mRNA level and left ventricular mass (LVM) indexed to body weight. The analysis included experimental groups after 8 and 10 weeks of aortic constriction, 8 week’s sham-operated and intact animals, r indicates Pearson coefficient; for all ∗ P
    Mouse Monoclonal Beta Sarcoglycan Antibodies, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 88/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals β sarcoglycan
    Impaired sarcolemmal glycoprotein trafficking in hearts overexpressing secretion-defective Thbs4. (A) Western blotting for membrane glycoproteins purified from hearts of transgenic mice overexpressing wild-type Thbs4 (DTG-Thbs4) or the secretion-defective mutant (DTG-mCa 2+ ). Cadherin was used as a loading control. (B) Representative experiment of immunocytochemistry for <t>β-sarcoglycan</t> and the Golgi marker GM130 in cardiomyocytes isolated from transgenic hearts of the indicated genotype. Scale bar = 50 μm. Similar results were obtained in 2 additional experiments.
    β Sarcoglycan, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 87/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam δ sarcoglycan
    Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, <t>δ-sarcoglycan;</t> α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P
    δ Sarcoglycan, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Abcam anti δ sarcoglycan leica
    Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, <t>δ-sarcoglycan;</t> α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P
    Anti δ Sarcoglycan Leica, supplied by Abcam, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals rabbit polyclonal anti β sarcoglycan
    Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, <t>δ-sarcoglycan;</t> α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P
    Rabbit Polyclonal Anti β Sarcoglycan, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    R&D Systems goat anti human δ sarcoglycan
    Adenoviral gene transfer efficiency and expression of wild-type (WT) and mutant <t>δ-sarcoglycan</t> (δSG) in adult cardiac myocytes. A : δSG adenoviral vector design. B : time course of δSG expression using adenoviral vectors in
    Goat Anti Human δ Sarcoglycan, supplied by R&D Systems, used in various techniques. Bioz Stars score: 83/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Abcam anti human specific γ sarcoglycan antibody
    Adenoviral gene transfer efficiency and expression of wild-type (WT) and mutant <t>δ-sarcoglycan</t> (δSG) in adult cardiac myocytes. A : δSG adenoviral vector design. B : time course of δSG expression using adenoviral vectors in
    Anti Human Specific γ Sarcoglycan Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression of S151A-SGCD precedes cardiac function changes in adult Drosophila expressing S151A-SGCD. (A).Temperature shift from 18°C to 26°C results in deterioration in cardiac function and induction of S151A-SGCD expression. After a temperature shift to 26°C, the induction of transgene expression precedes the development of dilated cardiomyopathy by 48 hours as demonstrated by QRT-PCR measurements of S151A-SGCD mRNA levels compared to cardiac function by serial OCT. A second temperature shift back to 18°C represses S151A-SGCD mRNA expression and results in restoration of cardiac function by 48 hours. We performed three independent experiments using different batches of flies each time. The summary data for QRT-PCR at the indicated times and temperatures are expressed as the mean +/− SE of three independent experiments, each performed in triplicate. * p

    Journal: PLoS ONE

    Article Title: Serial Examination of an Inducible and Reversible Dilated Cardiomyopathy in Individual Adult Drosophila

    doi: 10.1371/journal.pone.0007132

    Figure Lengend Snippet: Expression of S151A-SGCD precedes cardiac function changes in adult Drosophila expressing S151A-SGCD. (A).Temperature shift from 18°C to 26°C results in deterioration in cardiac function and induction of S151A-SGCD expression. After a temperature shift to 26°C, the induction of transgene expression precedes the development of dilated cardiomyopathy by 48 hours as demonstrated by QRT-PCR measurements of S151A-SGCD mRNA levels compared to cardiac function by serial OCT. A second temperature shift back to 18°C represses S151A-SGCD mRNA expression and results in restoration of cardiac function by 48 hours. We performed three independent experiments using different batches of flies each time. The summary data for QRT-PCR at the indicated times and temperatures are expressed as the mean +/− SE of three independent experiments, each performed in triplicate. * p

    Article Snippet: Applied Biosystems Taqman Gene expression assays were used to perform quantitative (real time) RT-PCR (human delta-sarcoglycan, Hs01087180_m1 and fly ribosomal protein L32, Dm02151827_g1 for endogenous control).

    Techniques: Expressing, Quantitative RT-PCR

    Expression of human mutant S151A-SGCD causes an inducible and reversible dilated cardiomyopathy in adult Drosophila . (A) Temperature shift from 18°C to 26°C causes the induction of S151A-SGCD expression and subsequent deterioration in cardiac function. At 96 hours post induction, flies expressing S151A-SGCD demonstrate an enlargement in EDD and ESD with a resultant impairment in FS. At 96 hours, a temperature shift back to 18°C results in a repression of S151A-SGCD expression and subsequent improvement in cardiac function with return of EDD, ESD and FS to near baseline. A similar level of wt-SGCD expression after temperature shift from 18°C to 26°C does not result in deterioration in cardiac function. Each graph represents the summary data for serial OCT measurements of EDD, ESD and FS and are expressed as the mean +/− SE (n = 16 for wt-SGCD and n = 16 for S151A-SGCD). * P

    Journal: PLoS ONE

    Article Title: Serial Examination of an Inducible and Reversible Dilated Cardiomyopathy in Individual Adult Drosophila

    doi: 10.1371/journal.pone.0007132

    Figure Lengend Snippet: Expression of human mutant S151A-SGCD causes an inducible and reversible dilated cardiomyopathy in adult Drosophila . (A) Temperature shift from 18°C to 26°C causes the induction of S151A-SGCD expression and subsequent deterioration in cardiac function. At 96 hours post induction, flies expressing S151A-SGCD demonstrate an enlargement in EDD and ESD with a resultant impairment in FS. At 96 hours, a temperature shift back to 18°C results in a repression of S151A-SGCD expression and subsequent improvement in cardiac function with return of EDD, ESD and FS to near baseline. A similar level of wt-SGCD expression after temperature shift from 18°C to 26°C does not result in deterioration in cardiac function. Each graph represents the summary data for serial OCT measurements of EDD, ESD and FS and are expressed as the mean +/− SE (n = 16 for wt-SGCD and n = 16 for S151A-SGCD). * P

    Article Snippet: Applied Biosystems Taqman Gene expression assays were used to perform quantitative (real time) RT-PCR (human delta-sarcoglycan, Hs01087180_m1 and fly ribosomal protein L32, Dm02151827_g1 for endogenous control).

    Techniques: Expressing, Mutagenesis

    Isolation of alpha-sarcoglycan + EVs from plasma using immunoaffinity capturing. Anti-alpha-sarcoglycan antibodies were conjugated to magnetic beads to isolate muscle EVs from plasma. Western blot analysis confirmed the presence of the exosomal marker Tsg101 in isolated EVs. Ponceau S Staining has been used as loading control (a). MicroRNA quantifications showed an increase in the miR-206/miR-16 ratio in the SGCA + sub-population of EVs (SGCA-Beads) compared to total (total plasma EVs) or uncaptured EVs (Supernatant). MiR-206 expression levels were normalized versus the endogenous reference miR-16 and expressed as -ΔCq (where ΔCq = Cq miR-206 -Cq miR-16 ) (b). Moreover, the quantification of miR-16 ratio in the SGCA + sub-population of EVs compared to total or uncaptured EVs shows that SGCA-conjugated beads retained about 2–5% of the total amount of EVs, miR-16 expression levels were normalized versus the spike-in reference cel-miR-39 and expressed as -ΔCq (where ΔCq = Cq miR-16 -Cq cel-miR-39 ) (c). Asterisks denote significant changes ( p

    Journal: PLoS ONE

    Article Title: Muscle Releases Alpha-Sarcoglycan Positive Extracellular Vesicles Carrying miRNAs in the Bloodstream

    doi: 10.1371/journal.pone.0125094

    Figure Lengend Snippet: Isolation of alpha-sarcoglycan + EVs from plasma using immunoaffinity capturing. Anti-alpha-sarcoglycan antibodies were conjugated to magnetic beads to isolate muscle EVs from plasma. Western blot analysis confirmed the presence of the exosomal marker Tsg101 in isolated EVs. Ponceau S Staining has been used as loading control (a). MicroRNA quantifications showed an increase in the miR-206/miR-16 ratio in the SGCA + sub-population of EVs (SGCA-Beads) compared to total (total plasma EVs) or uncaptured EVs (Supernatant). MiR-206 expression levels were normalized versus the endogenous reference miR-16 and expressed as -ΔCq (where ΔCq = Cq miR-206 -Cq miR-16 ) (b). Moreover, the quantification of miR-16 ratio in the SGCA + sub-population of EVs compared to total or uncaptured EVs shows that SGCA-conjugated beads retained about 2–5% of the total amount of EVs, miR-16 expression levels were normalized versus the spike-in reference cel-miR-39 and expressed as -ΔCq (where ΔCq = Cq miR-16 -Cq cel-miR-39 ) (c). Asterisks denote significant changes ( p

    Article Snippet: Isolation of exosomes using alpha-sarcoglycan immunoaffinity capture beads Exosome-Dynabeads Streptavidin (Life technologies) in PBS + 0.1% BSA were mixed with biotinylated anti-alpha-sarcoglycan (clone AD1/20A6 Monosan) according to the manufacturer’s instructions.

    Techniques: Isolation, Magnetic Beads, Western Blot, Marker, Staining, Expressing

    Cross-sectional profiles of regenerating fibres. Three serial frozen cross sections of a biopsy obtained from regenerating vastus lateralis skeletal muscle 7 days after injury induced by electrical stimulation-elicited eccentric contractions (right). Three serial cross sections from the control (uninjured) leg of the same individual are shown (left) for comparison. The sections have been stained with alpha-sarcoglycan, beta-dystroglycan or dystrophin to label the sarcolemma, along with a basement membrane protein (laminin or collagen IV) and a myogenic marker (desmin or neonatal/embryonic myosin; MHCn/e). Each column of images contains single channel and combined images for each staining. In the injured muscle, dystrophin staining is completely absent in several fibres, while the basement membrane (laminin) is preserved. MHCn/e staining is evident in some small dystrophin-negative fibres. A similar pattern is evident from the alpha-sarcoglycan and beta-dystroglycan staining, with negative fibres, together with desmin+ cells, contained within a preserved basement membrane (collagen IV). Asterisk indicates some of the necrotic fibres. Note the different profiles of the injured fibres, such as varying fibre size, and infiltrating cells either confined to the fibre periphery or dispersed throughout the fibre. Scale bar, 100 μm

    Journal: Skeletal Muscle

    Article Title: The breaking and making of healthy adult human skeletal muscle in vivo

    doi: 10.1186/s13395-017-0142-x

    Figure Lengend Snippet: Cross-sectional profiles of regenerating fibres. Three serial frozen cross sections of a biopsy obtained from regenerating vastus lateralis skeletal muscle 7 days after injury induced by electrical stimulation-elicited eccentric contractions (right). Three serial cross sections from the control (uninjured) leg of the same individual are shown (left) for comparison. The sections have been stained with alpha-sarcoglycan, beta-dystroglycan or dystrophin to label the sarcolemma, along with a basement membrane protein (laminin or collagen IV) and a myogenic marker (desmin or neonatal/embryonic myosin; MHCn/e). Each column of images contains single channel and combined images for each staining. In the injured muscle, dystrophin staining is completely absent in several fibres, while the basement membrane (laminin) is preserved. MHCn/e staining is evident in some small dystrophin-negative fibres. A similar pattern is evident from the alpha-sarcoglycan and beta-dystroglycan staining, with negative fibres, together with desmin+ cells, contained within a preserved basement membrane (collagen IV). Asterisk indicates some of the necrotic fibres. Note the different profiles of the injured fibres, such as varying fibre size, and infiltrating cells either confined to the fibre periphery or dispersed throughout the fibre. Scale bar, 100 μm

    Article Snippet: Sections were stained with various combinations of antibodies against laminin, CD56, desmin and embryonic myosin (F1.652; Developmental Studies Hybridoma Bank); neonatal myosin (NCL-MHCn; Novocastra, Leica Microsystems A/S, Ballerup, Denmark); alpha-sarcoglycan (NCL-L-a-SARC, Novocastra); beta-dystroglycan (NCL-L-a-SARC, Novocastra); myogenin (F5d, Developmental Studies Hybridoma Bank); nestin, CD68, collagen IV and dystrophin (cat. no. D8168, Sigma-Aldrich Denmark A/S, Copenhagen, Denmark); myosin type I (BA.D5, Developmental Studies Hybridoma Bank) and myosin type II (A4.74, Developmental Studies Hybridoma Bank).

    Techniques: Staining, Marker

    Ameliorated NMJ fragmentation and denervation in aged mice by SGα overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.

    Journal: The Journal of Neuroscience

    Article Title: Sarcoglycan Alpha Mitigates Neuromuscular Junction Decline in Aged Mice by Stabilizing LRP4

    doi: 10.1523/JNEUROSCI.0860-18.2018

    Figure Lengend Snippet: Ameliorated NMJ fragmentation and denervation in aged mice by SGα overexpression. A , Reduced fragmentation and improved innervation. Shown were TA muscles of mice at different ages and indicated AAV treatment, whole-mount stained with CF568 α-BTX (red) and anti-NF/Syn antibodies (visualized by AlexaFluor 488 goat anti-rabbit IgG; green). Blue arrowhead, denervated or partially innervated endplate. Scale bar, 50 μm. B , Enlarged images of individual endplates. Scale bar, 10 μm. C – F , Quantitative analysis of data in A . C , Rescue of denervation in aged mice by SGα expression; F (2,12) = 29, *** p = 0.0001, ** p = 0.0051. D , Rescue of fragmentation number in aged mice by SGα expression; F (2,12) = 25.8, *** p = 0.0001, ** p = 0.0023. E , Rescue of fragmented endplate percentage in aged mice by SGα expression; F (2,12) = 93, *** p = 0.0001, *** p = 0.0003. F , Rescue of reduced AChR cluster intensity in aged mice by SGα expression; F (2,12) = 35.9, *** p = 0.0001, ** p = 0.0028; N = 5 mice per group, one-way ANOVA.

    Article Snippet: Antibodies used were as follows: AChRδ (88B; 1:2000 for Western blot) from ThermoFisher Scientific; AChRε (ab65180; 1:2000 for Western blot), SGα (ab189254; 1:500 for staining and 1:2000 for Western blot) from Abcam; Ubiquitin (sc-8017; 1:1000 for Western blot), AChRα (sc-65829; 1:1000 for Western blot), and AChRβ (sc-11371; 1:1000 for Western blot) from Santa Cruz Biotechnology; DOK7 (AF6398; 1:1000 for Western blot) from R & D Systems; neurofilament (C28E10; 1:500 for staining) and synapsin (D12G5, 1:500 for staining) from Cell Signaling Technology; GAPDH (NB 600-501; 1:3000 for Western blot) from Novus; V5 (V8012; 1:1000 for Western blot), laminin (041M4799; 1:200 for staining) and GFP (11814460001; 1:3000 for Western blot and 1:500 for staining) from Sigma Aldrich.

    Techniques: Mouse Assay, Over Expression, Staining, Expressing

    Restored muscle size and strength in aged mice by SGα expression. A , Increased cross-section area and reduced central nuclei in aged muscle by SGα expression. Cross-sections of TA muscle were stained with laminin antibody (red) and DAPI (blue). Yellow arrowhead, central nuclei. Scale bar, 20 μm. B , Increased cross-section area in aged mice by SGα expression; F (2,9) = 24.1, *** p = 0.0003, ** p = 0.0012. C , Reduced central nuclei in aged mice by SGα expression; F (2,9) = 27.8, *** p = 0.0002, *** p = 0.0006; N = 4 mice per group, one-way ANOVA. D , Representative twitch (top) and tetanic force at 50 and 150 Hz (bottom) by sciatic nerve stimulation. E , Rescue of reduced twitch force in aged mice by SGα expression; F (2,9) = 35.1, *** p = 0.0001 and 0.0008. F , Rescue of reduced tetanic force in aged mice by SGα expression; F (2,36) = 34, * p

    Journal: The Journal of Neuroscience

    Article Title: Sarcoglycan Alpha Mitigates Neuromuscular Junction Decline in Aged Mice by Stabilizing LRP4

    doi: 10.1523/JNEUROSCI.0860-18.2018

    Figure Lengend Snippet: Restored muscle size and strength in aged mice by SGα expression. A , Increased cross-section area and reduced central nuclei in aged muscle by SGα expression. Cross-sections of TA muscle were stained with laminin antibody (red) and DAPI (blue). Yellow arrowhead, central nuclei. Scale bar, 20 μm. B , Increased cross-section area in aged mice by SGα expression; F (2,9) = 24.1, *** p = 0.0003, ** p = 0.0012. C , Reduced central nuclei in aged mice by SGα expression; F (2,9) = 27.8, *** p = 0.0002, *** p = 0.0006; N = 4 mice per group, one-way ANOVA. D , Representative twitch (top) and tetanic force at 50 and 150 Hz (bottom) by sciatic nerve stimulation. E , Rescue of reduced twitch force in aged mice by SGα expression; F (2,9) = 35.1, *** p = 0.0001 and 0.0008. F , Rescue of reduced tetanic force in aged mice by SGα expression; F (2,36) = 34, * p

    Article Snippet: Antibodies used were as follows: AChRδ (88B; 1:2000 for Western blot) from ThermoFisher Scientific; AChRε (ab65180; 1:2000 for Western blot), SGα (ab189254; 1:500 for staining and 1:2000 for Western blot) from Abcam; Ubiquitin (sc-8017; 1:1000 for Western blot), AChRα (sc-65829; 1:1000 for Western blot), and AChRβ (sc-11371; 1:1000 for Western blot) from Santa Cruz Biotechnology; DOK7 (AF6398; 1:1000 for Western blot) from R & D Systems; neurofilament (C28E10; 1:500 for staining) and synapsin (D12G5, 1:500 for staining) from Cell Signaling Technology; GAPDH (NB 600-501; 1:3000 for Western blot) from Novus; V5 (V8012; 1:1000 for Western blot), laminin (041M4799; 1:200 for staining) and GFP (11814460001; 1:3000 for Western blot and 1:500 for staining) from Sigma Aldrich.

    Techniques: Mouse Assay, Expressing, Staining

    Increasing LRP4 protein stability by SGα. A , LRP4 interaction with SGα, but not SGδ. Flag-LRP4 was precipitated from HEK293 cells cotransfected with SGα-V5 or SGδ-V5 and probed with anti-V5 antibody. Lysates were probed with the same antibodies as control. B , In vivo interaction between LRP4 and SGα. Homogenates were prepared from muscles of Flag-Lrp4 transgenic mice and subjected to IP with anti-SGα antibody. Immunocomplex was probed with anti-Flag and SGα antibodies. C , mRNA expression of sarcoglycan complex molecules in diaphragm synaptic region; t (4) = 9.97, ** p = 0.0026 for Sgca ; t (4) = 8.87, ** p = 0.0039 for Sgcb ; t (4) = 10.8, *** p = 0.0004 for Sgcg ; N = 3 mice per group, unpaired t test. D , Reduced SGα protein level in aged mice. Top, Representative blots. Bottom, Quantitative data; t (4) = 5.66, * p = 0.015; N = 3 mice per group, unpaired t test. E , Discontinuous staining of SGα in aged TA muscles. White arrowhead, NMJs identified by α-BTX with discontinuous SGα staining. Scale bar, 20 μm. F , G , Increased LRP4 stability in HEK293 cells expressing SGα. HEK293 cells were transfected with Flag-LRP4 without (control) or with SGα-V5 and incubated with CHX. Flag-LRP4 levels were analyzed at indicated times. F , Representative blot; G , quantitative data; F (1,16) = 56.5, ** p = 0.0021 and 0.0095, *** p = 0.0001; N = 3, two-way ANOVA.

    Journal: The Journal of Neuroscience

    Article Title: Sarcoglycan Alpha Mitigates Neuromuscular Junction Decline in Aged Mice by Stabilizing LRP4

    doi: 10.1523/JNEUROSCI.0860-18.2018

    Figure Lengend Snippet: Increasing LRP4 protein stability by SGα. A , LRP4 interaction with SGα, but not SGδ. Flag-LRP4 was precipitated from HEK293 cells cotransfected with SGα-V5 or SGδ-V5 and probed with anti-V5 antibody. Lysates were probed with the same antibodies as control. B , In vivo interaction between LRP4 and SGα. Homogenates were prepared from muscles of Flag-Lrp4 transgenic mice and subjected to IP with anti-SGα antibody. Immunocomplex was probed with anti-Flag and SGα antibodies. C , mRNA expression of sarcoglycan complex molecules in diaphragm synaptic region; t (4) = 9.97, ** p = 0.0026 for Sgca ; t (4) = 8.87, ** p = 0.0039 for Sgcb ; t (4) = 10.8, *** p = 0.0004 for Sgcg ; N = 3 mice per group, unpaired t test. D , Reduced SGα protein level in aged mice. Top, Representative blots. Bottom, Quantitative data; t (4) = 5.66, * p = 0.015; N = 3 mice per group, unpaired t test. E , Discontinuous staining of SGα in aged TA muscles. White arrowhead, NMJs identified by α-BTX with discontinuous SGα staining. Scale bar, 20 μm. F , G , Increased LRP4 stability in HEK293 cells expressing SGα. HEK293 cells were transfected with Flag-LRP4 without (control) or with SGα-V5 and incubated with CHX. Flag-LRP4 levels were analyzed at indicated times. F , Representative blot; G , quantitative data; F (1,16) = 56.5, ** p = 0.0021 and 0.0095, *** p = 0.0001; N = 3, two-way ANOVA.

    Article Snippet: Antibodies used were as follows: AChRδ (88B; 1:2000 for Western blot) from ThermoFisher Scientific; AChRε (ab65180; 1:2000 for Western blot), SGα (ab189254; 1:500 for staining and 1:2000 for Western blot) from Abcam; Ubiquitin (sc-8017; 1:1000 for Western blot), AChRα (sc-65829; 1:1000 for Western blot), and AChRβ (sc-11371; 1:1000 for Western blot) from Santa Cruz Biotechnology; DOK7 (AF6398; 1:1000 for Western blot) from R & D Systems; neurofilament (C28E10; 1:500 for staining) and synapsin (D12G5, 1:500 for staining) from Cell Signaling Technology; GAPDH (NB 600-501; 1:3000 for Western blot) from Novus; V5 (V8012; 1:1000 for Western blot), laminin (041M4799; 1:200 for staining) and GFP (11814460001; 1:3000 for Western blot and 1:500 for staining) from Sigma Aldrich.

    Techniques: In Vivo, Transgenic Assay, Mouse Assay, Expressing, Staining, Transfection, Incubation

    Increased LRP4 level with reduced ubiquitination in aged muscles by SGα expression. A , Diagram of AAV9-SGα-GFP construct. Human SGCA was inserted at N-terminus of GFP as a fusion protein. ITR, Inverted terminal repeats; CMV, cytomegalovirus promoter; β-Glob, β-globulin intron. B , SGα expression in HEK293 cells transfected with AAV9-SGα-GFP. C , Expression of GFP in muscles after intramuscular and intravenous injection with indicated AAV. Shown were TA cross-sections. D , High infection rates by AAV9-expressing SGα-GFP. E , Increased protein level and reduced ubiquitination of LRP4 by SGα expression. Muscles were isolated 6 weeks after intravenous injection of AAV9-expressing SGα. F , G , Quantification of data in E ; F (2,6) = 16.7, ** p = 0.0029, * p = 0.035 for F ; F (2,6) = 29.7, ** p = 0.0021, *** p = 0.0009 for G ; N = 3 mice per group, one-way ANOVA.

    Journal: The Journal of Neuroscience

    Article Title: Sarcoglycan Alpha Mitigates Neuromuscular Junction Decline in Aged Mice by Stabilizing LRP4

    doi: 10.1523/JNEUROSCI.0860-18.2018

    Figure Lengend Snippet: Increased LRP4 level with reduced ubiquitination in aged muscles by SGα expression. A , Diagram of AAV9-SGα-GFP construct. Human SGCA was inserted at N-terminus of GFP as a fusion protein. ITR, Inverted terminal repeats; CMV, cytomegalovirus promoter; β-Glob, β-globulin intron. B , SGα expression in HEK293 cells transfected with AAV9-SGα-GFP. C , Expression of GFP in muscles after intramuscular and intravenous injection with indicated AAV. Shown were TA cross-sections. D , High infection rates by AAV9-expressing SGα-GFP. E , Increased protein level and reduced ubiquitination of LRP4 by SGα expression. Muscles were isolated 6 weeks after intravenous injection of AAV9-expressing SGα. F , G , Quantification of data in E ; F (2,6) = 16.7, ** p = 0.0029, * p = 0.035 for F ; F (2,6) = 29.7, ** p = 0.0021, *** p = 0.0009 for G ; N = 3 mice per group, one-way ANOVA.

    Article Snippet: Antibodies used were as follows: AChRδ (88B; 1:2000 for Western blot) from ThermoFisher Scientific; AChRε (ab65180; 1:2000 for Western blot), SGα (ab189254; 1:500 for staining and 1:2000 for Western blot) from Abcam; Ubiquitin (sc-8017; 1:1000 for Western blot), AChRα (sc-65829; 1:1000 for Western blot), and AChRβ (sc-11371; 1:1000 for Western blot) from Santa Cruz Biotechnology; DOK7 (AF6398; 1:1000 for Western blot) from R & D Systems; neurofilament (C28E10; 1:500 for staining) and synapsin (D12G5, 1:500 for staining) from Cell Signaling Technology; GAPDH (NB 600-501; 1:3000 for Western blot) from Novus; V5 (V8012; 1:1000 for Western blot), laminin (041M4799; 1:200 for staining) and GFP (11814460001; 1:3000 for Western blot and 1:500 for staining) from Sigma Aldrich.

    Techniques: Expressing, Construct, Transfection, Injection, Infection, Isolation, Mouse Assay

    Improved neuromuscular transmission in aged mice by SGα expression. A , B , Reduced ratio of 2nd–10th CMAP amplitudes over the first CMAP amplitude at 40 Hz stimulations. A , Representative traces; B , quantitative data; F (2,90) = 47.7, * p

    Journal: The Journal of Neuroscience

    Article Title: Sarcoglycan Alpha Mitigates Neuromuscular Junction Decline in Aged Mice by Stabilizing LRP4

    doi: 10.1523/JNEUROSCI.0860-18.2018

    Figure Lengend Snippet: Improved neuromuscular transmission in aged mice by SGα expression. A , B , Reduced ratio of 2nd–10th CMAP amplitudes over the first CMAP amplitude at 40 Hz stimulations. A , Representative traces; B , quantitative data; F (2,90) = 47.7, * p

    Article Snippet: Antibodies used were as follows: AChRδ (88B; 1:2000 for Western blot) from ThermoFisher Scientific; AChRε (ab65180; 1:2000 for Western blot), SGα (ab189254; 1:500 for staining and 1:2000 for Western blot) from Abcam; Ubiquitin (sc-8017; 1:1000 for Western blot), AChRα (sc-65829; 1:1000 for Western blot), and AChRβ (sc-11371; 1:1000 for Western blot) from Santa Cruz Biotechnology; DOK7 (AF6398; 1:1000 for Western blot) from R & D Systems; neurofilament (C28E10; 1:500 for staining) and synapsin (D12G5, 1:500 for staining) from Cell Signaling Technology; GAPDH (NB 600-501; 1:3000 for Western blot) from Novus; V5 (V8012; 1:1000 for Western blot), laminin (041M4799; 1:200 for staining) and GFP (11814460001; 1:3000 for Western blot and 1:500 for staining) from Sigma Aldrich.

    Techniques: Transmission Assay, Mouse Assay, Expressing

    Morphological and molecular evidences of myocardial hypertrophy progression. (A) Immunohistochemical staining of beta- sarcoglycan allowing to detect outer membrane showing clear increase of cells size in transverse orientation by 10-weeks group compared to intact. (B) Increase in cell diameter (Dmin) illustrating progressive cardiomyocytes enlargement, and significant increase after 8 and 10 weeks of aortic banding performing. Nppa expression was upregulated in LV (C) and IVS (D) after 1, 2, 8, and 10 weeks of model duration compared to intact or sham-control groups. (E) Positive linear correlation was found for Nppa mRNA level and left ventricular mass (LVM) indexed to body weight. The analysis included experimental groups after 8 and 10 weeks of aortic constriction, 8 week’s sham-operated and intact animals, r indicates Pearson coefficient; for all ∗ P

    Journal: Frontiers in Genetics

    Article Title: Time- and Ventricular-Specific Expression Profiles of Genes Encoding Z-Disk Proteins in Pressure Overload Model of Left Ventricular Hypertrophy

    doi: 10.3389/fgene.2018.00684

    Figure Lengend Snippet: Morphological and molecular evidences of myocardial hypertrophy progression. (A) Immunohistochemical staining of beta- sarcoglycan allowing to detect outer membrane showing clear increase of cells size in transverse orientation by 10-weeks group compared to intact. (B) Increase in cell diameter (Dmin) illustrating progressive cardiomyocytes enlargement, and significant increase after 8 and 10 weeks of aortic banding performing. Nppa expression was upregulated in LV (C) and IVS (D) after 1, 2, 8, and 10 weeks of model duration compared to intact or sham-control groups. (E) Positive linear correlation was found for Nppa mRNA level and left ventricular mass (LVM) indexed to body weight. The analysis included experimental groups after 8 and 10 weeks of aortic constriction, 8 week’s sham-operated and intact animals, r indicates Pearson coefficient; for all ∗ P

    Article Snippet: After blocking in 15% fetal calf serum (FCS) for 30 min at room temperature, samples were incubated overnight at +4°C with mouse monoclonal beta-sarcoglycan antibodies (Leica Biosystems, NCL-L-b-SARC).

    Techniques: Immunohistochemistry, Staining, Expressing

    Impaired sarcolemmal glycoprotein trafficking in hearts overexpressing secretion-defective Thbs4. (A) Western blotting for membrane glycoproteins purified from hearts of transgenic mice overexpressing wild-type Thbs4 (DTG-Thbs4) or the secretion-defective mutant (DTG-mCa 2+ ). Cadherin was used as a loading control. (B) Representative experiment of immunocytochemistry for β-sarcoglycan and the Golgi marker GM130 in cardiomyocytes isolated from transgenic hearts of the indicated genotype. Scale bar = 50 μm. Similar results were obtained in 2 additional experiments.

    Journal: Molecular and Cellular Biology

    Article Title: Defective Flux of Thrombospondin-4 through the Secretory Pathway Impairs Cardiomyocyte Membrane Stability and Causes Cardiomyopathy

    doi: 10.1128/MCB.00114-18

    Figure Lengend Snippet: Impaired sarcolemmal glycoprotein trafficking in hearts overexpressing secretion-defective Thbs4. (A) Western blotting for membrane glycoproteins purified from hearts of transgenic mice overexpressing wild-type Thbs4 (DTG-Thbs4) or the secretion-defective mutant (DTG-mCa 2+ ). Cadherin was used as a loading control. (B) Representative experiment of immunocytochemistry for β-sarcoglycan and the Golgi marker GM130 in cardiomyocytes isolated from transgenic hearts of the indicated genotype. Scale bar = 50 μm. Similar results were obtained in 2 additional experiments.

    Article Snippet: Proteins were resolved by SDS-PAGE, transferred to polyvinylidene difluoride (PVDF) membranes (Millipore), and immunoblotted with antibodies for Flag (Cell Signaling Technology or Sigma), calreticulin, PDI, integrin β1, pan-cadherin, and Rab6 (Cell Signaling Technology), Armet, SAR1, δ-sarcoglycan, and β-actin (Abcam), α-sarcoglycan and β-dystroglycan (Developmental Studies Hybridoma Bank), β-sarcoglycan (Novus Biologicals), Rab24 (BD Transduction), atrial natriuretic factor (ANF; Millipore), α-tubulin, BiP, and α-sarcomeric actin (Sigma), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Fitzgerald), and Thbs4 (Santa Cruz).

    Techniques: Western Blot, Purification, Transgenic Assay, Mouse Assay, Mutagenesis, Immunocytochemistry, Marker, Isolation

    Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, δ-sarcoglycan; α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P

    Journal: Human Molecular Genetics

    Article Title: Genetic overexpression of Serpina3n attenuates muscular dystrophy in mice

    doi: 10.1093/hmg/ddw005

    Figure Lengend Snippet: Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, δ-sarcoglycan; α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P

    Article Snippet: Primary antibodies used included: dystrophin [MANDYS1(3B7) 1:500]; utrophin (DSHB Mancho3-8A4 1:500); α7-integrin (Santa Cruz SC-27706 1:500); β1D-integrin (Chemicon; MAB1900, 1:200); δ-sarcoglycan (Abcam ab137101 1:1000); α-dystroglycan (Millipore 05-593 1:500); β-dystroglycan (MANDAG2 clone 7D11 1:500); β-sarcoglycan (Vector labs VP-B206 1: 500); α-sarcoglycan (DSHB IVD3(1)A9 1:500) and CaV1.1 (Pierce MA3-920 1:1000).

    Techniques: Over Expression, Immunohistochemistry, Western Blot, Mouse Assay, Staining

    Adenoviral gene transfer efficiency and expression of wild-type (WT) and mutant δ-sarcoglycan (δSG) in adult cardiac myocytes. A : δSG adenoviral vector design. B : time course of δSG expression using adenoviral vectors in

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Dilated cardiomyopathy mutations in δ-sarcoglycan exert a dominant-negative effect on cardiac myocyte mechanical stability

    doi: 10.1152/ajpheart.00521.2015

    Figure Lengend Snippet: Adenoviral gene transfer efficiency and expression of wild-type (WT) and mutant δ-sarcoglycan (δSG) in adult cardiac myocytes. A : δSG adenoviral vector design. B : time course of δSG expression using adenoviral vectors in

    Article Snippet: Primary antibodies included: A-14 rabbit polyclonal anti-myc (Santa Cruz Biotechnology; 1:100), goat anti-human δ-sarcoglycan (R & D Systems; 1:1,000), IIH6 anti-α dystroglycan (Millipore; 1:1,000), and mouse anti-β dystroglycan (Vector Laboratories; 1:1,000).

    Techniques: Expressing, Mutagenesis, Plasmid Preparation