anti α sarcoglycan mouse monoclonal antibody  (Novocastra)

 
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    Structured Review

    Novocastra anti α sarcoglycan mouse monoclonal antibody
    I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, <t>α-sarcoglycan,</t> and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.
    Anti α Sarcoglycan Mouse Monoclonal Antibody, supplied by Novocastra, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti α sarcoglycan mouse monoclonal antibody/product/Novocastra
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti α sarcoglycan mouse monoclonal antibody - by Bioz Stars, 2022-09
    85/100 stars

    Images

    1) Product Images from "Long-Term Efficacy of Systemic Multiexon Skipping Targeting Dystrophin Exons 45–55 With a Cocktail of Vivo-Morpholinos in Mdx52 Mice"

    Article Title: Long-Term Efficacy of Systemic Multiexon Skipping Targeting Dystrophin Exons 45–55 With a Cocktail of Vivo-Morpholinos in Mdx52 Mice

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1038/mtna.2014.76

    I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, α-sarcoglycan, and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.
    Figure Legend Snippet: I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, α-sarcoglycan, and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.

    Techniques Used: Staining, Mouse Assay

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    Novocastra anti α sarcoglycan mouse monoclonal antibody
    I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, <t>α-sarcoglycan,</t> and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.
    Anti α Sarcoglycan Mouse Monoclonal Antibody, supplied by Novocastra, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti α sarcoglycan mouse monoclonal antibody/product/Novocastra
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti α sarcoglycan mouse monoclonal antibody - by Bioz Stars, 2022-09
    85/100 stars
      Buy from Supplier

    90
    Novocastra monoclonal α sarcoglycan ab
    ) to create MHG or MHD, respectively. Transgenic mice with the MHG transgene were crossed with <t>γ-sarcoglycan–null</t> ( gsg –/– ) mice to generate gsg –/– /MHG mice. Transgenic mice with the MHD transgene were crossed with δ-sarcoglycan–null ( dsg –/– ) mice generating dsg –/– /MHD mice. pA, polyadenylation signals. (B) Immunoblot of whole heart extracts from normal, gsg –/– , dsg –/– , gsg –/– /MHG, and dsg –/– /MHD animals at 12 weeks of age using γ-sarcoglycan Ab showed γ-sarcoglycan expression was absent in the hearts of gsg –/– and dsg –/– animals, but was restored in transgenic animals. Quantitative Western blot analysis determined the level of γ-sarcoglycan expression in gsg –/– /MHG animals to be sevenfold above normal. (C) δ-Sarcoglycan expression was restored to normal levels by expression of the MHD transgene in dsg –/– /MHD hearts. Expression of δ-sarcoglycan from the MHD transgene also resulted in recovery of γ-sarcoglycan to normal levels (last lane). (D) Immunoblots for the remaining sarcoglycan subunits showed that <t>α-sarcoglycan</t> (ASG) expression is recovered in hearts with either γ- or δ-sarcoglycan transgene expression. β-Sarcoglycan protein (BSG) is increased to normal levels, and ζ-sarcoglycan protein (ZSG) levels are not significantly different in transgenic hearts. Loading control is shown for B, C, and D. Coom., Coomassie blue.
    Monoclonal α Sarcoglycan Ab, supplied by Novocastra, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal α sarcoglycan ab/product/Novocastra
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal α sarcoglycan ab - by Bioz Stars, 2022-09
    90/100 stars
      Buy from Supplier

    90
    Novocastra monoclonal anti α sarcoglycan
    Verapamil treatment does not improve expression of the remaining <t>sarcoglycan</t> at the plasma membrane. The major sarcoglycan complex of cardiomyocytes is α-, β-, γ-, and δ-sarcoglycan. Antibodies specific to <t>α-sarcoglycan,</t> β-sarcoglycan, and δ-sarcoglycan were used on normal control and γ-sarcoglycan-null mice (gsg−/−) that were treated and untreated with verapamil. Scale bar, 10 μm.
    Monoclonal Anti α Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti α sarcoglycan/product/Novocastra
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti α sarcoglycan - by Bioz Stars, 2022-09
    90/100 stars
      Buy from Supplier

    91
    Novocastra mouse monoclonal anti α sarcoglycan
    Dependence of ecto-nucleotidase activity of <t>α-sarcoglycan-expressing</t> HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P
    Mouse Monoclonal Anti α Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti α sarcoglycan/product/Novocastra
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti α sarcoglycan - by Bioz Stars, 2022-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, α-sarcoglycan, and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Long-Term Efficacy of Systemic Multiexon Skipping Targeting Dystrophin Exons 45–55 With a Cocktail of Vivo-Morpholinos in Mdx52 Mice

    doi: 10.1038/mtna.2014.76

    Figure Lengend Snippet: I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, α-sarcoglycan, and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.

    Article Snippet: In systemic treatment with the 10-vPMO cocktail, the diaphragm, biceps femoris, quadriceps, gastrocnemius, tibialis anterior, biceps brachii, triceps brachii, and heart muscles were examined 2 weeks after the final injection using anti-dystrophin (P7) antibody and antibodies against dystrophin-associated proteins: anti-α1-syntrophin rabbit polyclonal antibody (1:200, Abcam, Cambridge, UK), anti-nNOS rabbit polyclonal antibody (1:100, Invitrogen), anti-α-sarcoglycan mouse monoclonal antibody (1:10, Novocastra Laboratories), and anti-β-dystroglycan mouse monoclonal antibody (1:5, Novocastra Laboratories).

    Techniques: Staining, Mouse Assay

    ) to create MHG or MHD, respectively. Transgenic mice with the MHG transgene were crossed with γ-sarcoglycan–null ( gsg –/– ) mice to generate gsg –/– /MHG mice. Transgenic mice with the MHD transgene were crossed with δ-sarcoglycan–null ( dsg –/– ) mice generating dsg –/– /MHD mice. pA, polyadenylation signals. (B) Immunoblot of whole heart extracts from normal, gsg –/– , dsg –/– , gsg –/– /MHG, and dsg –/– /MHD animals at 12 weeks of age using γ-sarcoglycan Ab showed γ-sarcoglycan expression was absent in the hearts of gsg –/– and dsg –/– animals, but was restored in transgenic animals. Quantitative Western blot analysis determined the level of γ-sarcoglycan expression in gsg –/– /MHG animals to be sevenfold above normal. (C) δ-Sarcoglycan expression was restored to normal levels by expression of the MHD transgene in dsg –/– /MHD hearts. Expression of δ-sarcoglycan from the MHD transgene also resulted in recovery of γ-sarcoglycan to normal levels (last lane). (D) Immunoblots for the remaining sarcoglycan subunits showed that α-sarcoglycan (ASG) expression is recovered in hearts with either γ- or δ-sarcoglycan transgene expression. β-Sarcoglycan protein (BSG) is increased to normal levels, and ζ-sarcoglycan protein (ZSG) levels are not significantly different in transgenic hearts. Loading control is shown for B, C, and D. Coom., Coomassie blue.

    Journal: Journal of Clinical Investigation

    Article Title: Smooth muscle cell-extrinsic vascular spasm arises from cardiomyocyte degeneration in sarcoglycan-deficient cardiomyopathy

    doi: 10.1172/JCI200420410

    Figure Lengend Snippet: ) to create MHG or MHD, respectively. Transgenic mice with the MHG transgene were crossed with γ-sarcoglycan–null ( gsg –/– ) mice to generate gsg –/– /MHG mice. Transgenic mice with the MHD transgene were crossed with δ-sarcoglycan–null ( dsg –/– ) mice generating dsg –/– /MHD mice. pA, polyadenylation signals. (B) Immunoblot of whole heart extracts from normal, gsg –/– , dsg –/– , gsg –/– /MHG, and dsg –/– /MHD animals at 12 weeks of age using γ-sarcoglycan Ab showed γ-sarcoglycan expression was absent in the hearts of gsg –/– and dsg –/– animals, but was restored in transgenic animals. Quantitative Western blot analysis determined the level of γ-sarcoglycan expression in gsg –/– /MHG animals to be sevenfold above normal. (C) δ-Sarcoglycan expression was restored to normal levels by expression of the MHD transgene in dsg –/– /MHD hearts. Expression of δ-sarcoglycan from the MHD transgene also resulted in recovery of γ-sarcoglycan to normal levels (last lane). (D) Immunoblots for the remaining sarcoglycan subunits showed that α-sarcoglycan (ASG) expression is recovered in hearts with either γ- or δ-sarcoglycan transgene expression. β-Sarcoglycan protein (BSG) is increased to normal levels, and ζ-sarcoglycan protein (ZSG) levels are not significantly different in transgenic hearts. Loading control is shown for B, C, and D. Coom., Coomassie blue.

    Article Snippet: Membranes were blocked in 5% milk in Tris-buffered saline with 0.1% Tween 20 and incubated with polyclonal γ-sarcoglycan Ab (1:1,000) , polyclonal δ-sarcoglycan Ab (1:2,000) , polyclonal ζ-sarcoglycan Ab (1:1,000) , polyclonal β-sarcoglycan Ab (1:2,000) , or monoclonal α-sarcoglycan Ab (NCL-ASG, 1:200; Novocastra, Newcastle-Upon-Tyne, United Kingdom).

    Techniques: Transgenic Assay, Mouse Assay, Expressing, Western Blot

    Verapamil treatment does not improve expression of the remaining sarcoglycan at the plasma membrane. The major sarcoglycan complex of cardiomyocytes is α-, β-, γ-, and δ-sarcoglycan. Antibodies specific to α-sarcoglycan, β-sarcoglycan, and δ-sarcoglycan were used on normal control and γ-sarcoglycan-null mice (gsg−/−) that were treated and untreated with verapamil. Scale bar, 10 μm.

    Journal: The American Journal of Pathology

    Article Title: Secondary Coronary Artery Vasospasm Promotes Cardiomyopathy Progression

    doi:

    Figure Lengend Snippet: Verapamil treatment does not improve expression of the remaining sarcoglycan at the plasma membrane. The major sarcoglycan complex of cardiomyocytes is α-, β-, γ-, and δ-sarcoglycan. Antibodies specific to α-sarcoglycan, β-sarcoglycan, and δ-sarcoglycan were used on normal control and γ-sarcoglycan-null mice (gsg−/−) that were treated and untreated with verapamil. Scale bar, 10 μm.

    Article Snippet: Cryosections of 7 to 10 μm thickness were incubated with polyclonal anti-γ-sarcoglycan (1:1000), polyclonal anti-δ-sarcoglycan (1:500), polyclonal anti-β-sarcoglycan (1:150), affinity-purified polyclonal anti-ε-sarcoglycan (1:50), affinity-purified polyclonal anti-ζ-sarcoglycan (1:500), polyclonal anti-dystrophin (1:5000), monoclonal anti-dystrophin (NCL-Dys2, 1:200; Novocastra Laboratories, Newcastle on Tyne, UK), monoclonal anti-α-sarcoglycan (NCL-asarc, 1:100; Novocastra), or monoclonal anti-smooth muscle α-actin (clone 1A4, 1:1000; Sigma, St. Louis, MO).

    Techniques: Expressing, Mouse Assay

    Dependence of ecto-nucleotidase activity of α-sarcoglycan-expressing HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Dependence of ecto-nucleotidase activity of α-sarcoglycan-expressing HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Activity Assay, Expressing, Stable Transfection, Transfection, Plasmid Preparation

    Transient transfection of HEK-293 cells with α-sarcoglycan ( A ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (α-SG), empty vector (pcDNA3) transfected cells and untransfected cells (wild-type) by using an antibody specific to α-sarcoglycan. ( B ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (HEK α-SG) either untreated (left two lanes) or treated with N -glycosidase F (right two lanes). Amounts of protein from HEK-293 cell lysates were estimated ( A , B ) by using an antibody against β-actin. ( C ) Phase-contrast image (left panel) and immunofluorescence staining of non-permeabilized α-sarcoglycan transiently transfected HEK-293 cells with the antibody directed against the extracellular domain of α-sarcoglycan (right panel). ( D ) Extracellular ATP hydrolysis of wild-type HEK-293 cells (wt), or cells transiently transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from three experiments performed in triplicate; ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Transient transfection of HEK-293 cells with α-sarcoglycan ( A ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (α-SG), empty vector (pcDNA3) transfected cells and untransfected cells (wild-type) by using an antibody specific to α-sarcoglycan. ( B ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (HEK α-SG) either untreated (left two lanes) or treated with N -glycosidase F (right two lanes). Amounts of protein from HEK-293 cell lysates were estimated ( A , B ) by using an antibody against β-actin. ( C ) Phase-contrast image (left panel) and immunofluorescence staining of non-permeabilized α-sarcoglycan transiently transfected HEK-293 cells with the antibody directed against the extracellular domain of α-sarcoglycan (right panel). ( D ) Extracellular ATP hydrolysis of wild-type HEK-293 cells (wt), or cells transiently transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from three experiments performed in triplicate; ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Transfection, Western Blot, Plasmid Preparation, Immunofluorescence, Staining

    Ecto-nucleotidase activity of HEK-293 cells stably transfected with α-sarcoglycan ( A ) ATP-hydrolysing activity, measured as indicated in the Experimental section, was performed in HEK-293 cells transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from four experiments performed in triplicate; ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Ecto-nucleotidase activity of HEK-293 cells stably transfected with α-sarcoglycan ( A ) ATP-hydrolysing activity, measured as indicated in the Experimental section, was performed in HEK-293 cells transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from four experiments performed in triplicate; ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Activity Assay, Stable Transfection, Transfection, Plasmid Preparation

    Effects of antibodies against α-sarcoglycan on the ecto-nucleotidase activity of HEK-293 cells stably expressing the protein ATP-hydrolysing activity of cells incubated in the absence or in the presence of a monoclonal antibody against the extracellular domain of α-sarcoglycan encompassing the ATP-binding site (+mAb) or a polyclonal antibody specific for the C-terminal domain of the protein (+pAb). HEK-293 cells were preincubated for 30 min at 4 °C with or without antibodies. Results are from four experiments performed in triplicate; ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Effects of antibodies against α-sarcoglycan on the ecto-nucleotidase activity of HEK-293 cells stably expressing the protein ATP-hydrolysing activity of cells incubated in the absence or in the presence of a monoclonal antibody against the extracellular domain of α-sarcoglycan encompassing the ATP-binding site (+mAb) or a polyclonal antibody specific for the C-terminal domain of the protein (+pAb). HEK-293 cells were preincubated for 30 min at 4 °C with or without antibodies. Results are from four experiments performed in triplicate; ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Activity Assay, Stable Transfection, Expressing, Incubation, Binding Assay

    Western-blot analysis and total ecto-nucleotidase activity of C2C12 cells ( A ) Western-blot analysis of cell lysates obtained from proliferating (P), confluent (C) and myotubes at different days of culture. Lysate proteins (50 μg) were separated on 10% SDS/polyacrylamide slab gels, blotted on nitrocellulose filters and probed with specific antibodies, as indicated. ( B ) ATP-hydrolysing activity of C2C12 myoblasts (Mb) and 4- and 8-day-old myotubes (4d-Mt and 8d-Mt respectively) was determined by measuring P i ]. Cells were incubated for 15 min in a medium with the following composition: 20 mM Hepes, 140 mM NaCl, 4 mM MgCl 2 and 2 mM CaCl 2 . The reaction was started by adding 4 mM ATP and terminated by withdrawing a supernatant aliquot. Results are from four experiments performed in triplicate. ( C ) Effect of polyclonal (+pAb) and monoclonal (+mAb) antibodies specific for α-sarcoglycan on total ecto-ATPase activity of 7-day-old C2C12 myotubes incubated 30 min as above. Results are from three experiments performed in triplicate. * P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Western-blot analysis and total ecto-nucleotidase activity of C2C12 cells ( A ) Western-blot analysis of cell lysates obtained from proliferating (P), confluent (C) and myotubes at different days of culture. Lysate proteins (50 μg) were separated on 10% SDS/polyacrylamide slab gels, blotted on nitrocellulose filters and probed with specific antibodies, as indicated. ( B ) ATP-hydrolysing activity of C2C12 myoblasts (Mb) and 4- and 8-day-old myotubes (4d-Mt and 8d-Mt respectively) was determined by measuring P i ]. Cells were incubated for 15 min in a medium with the following composition: 20 mM Hepes, 140 mM NaCl, 4 mM MgCl 2 and 2 mM CaCl 2 . The reaction was started by adding 4 mM ATP and terminated by withdrawing a supernatant aliquot. Results are from four experiments performed in triplicate. ( C ) Effect of polyclonal (+pAb) and monoclonal (+mAb) antibodies specific for α-sarcoglycan on total ecto-ATPase activity of 7-day-old C2C12 myotubes incubated 30 min as above. Results are from three experiments performed in triplicate. * P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Western Blot, Activity Assay, Incubation