antibodies anti na v 1 5  (Alomone Labs)


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    Structured Review

    Alomone Labs antibodies anti na v 1 5
    The primers sequences for real-time quantitative PCR.
    Antibodies Anti Na V 1 5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    antibodies anti na v 1 5 - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "Transcription factor Meis1 act as a new regulator of ischemic arrhythmias in mice"

    Article Title: Transcription factor Meis1 act as a new regulator of ischemic arrhythmias in mice

    Journal: Journal of Advanced Research

    doi: 10.1016/j.jare.2021.11.004


    Figure Legend Snippet: The primers sequences for real-time quantitative PCR.

    Techniques Used:

    Meis1 inhibits the down-regulation of SCN5A /Na V 1.5 expression in MI hearts and hypoxic cardiomyocytes. (A-B) The mRNA and protein expression of sodium channel subunit SCN5A /Na V 1.5 in the border zone of infracted hearts from Sham, MI + AAV9-NC and MI + AAV9-Meis1 group (n = 6). (C-D) The expression of SCN5A /Na V 1.5 at mRNA and protein level in neonatal mouse cardiomyocytes after treatment with hypoxia for 24 h with or without forced expression of Meis1 (n = 6). (E) Immunofluorescence staining for Na V 1.5 (green), α-actinin (red) proteins in neonatal mouse cardiomyocytes after treatment with hypoxia for 24 h with or without overexpression of Meis1 (left panels) and enlargement of outlined squares (right panels). Scale bar indicates 5 μm/2 μm. One-way ANOVA was used to determine statistical significance in these experiments. Error bars represent mean ± SEM of each group. * P < 0.05, ** P < 0.01. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: Meis1 inhibits the down-regulation of SCN5A /Na V 1.5 expression in MI hearts and hypoxic cardiomyocytes. (A-B) The mRNA and protein expression of sodium channel subunit SCN5A /Na V 1.5 in the border zone of infracted hearts from Sham, MI + AAV9-NC and MI + AAV9-Meis1 group (n = 6). (C-D) The expression of SCN5A /Na V 1.5 at mRNA and protein level in neonatal mouse cardiomyocytes after treatment with hypoxia for 24 h with or without forced expression of Meis1 (n = 6). (E) Immunofluorescence staining for Na V 1.5 (green), α-actinin (red) proteins in neonatal mouse cardiomyocytes after treatment with hypoxia for 24 h with or without overexpression of Meis1 (left panels) and enlargement of outlined squares (right panels). Scale bar indicates 5 μm/2 μm. One-way ANOVA was used to determine statistical significance in these experiments. Error bars represent mean ± SEM of each group. * P < 0.05, ** P < 0.01. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: Expressing, Immunofluorescence, Staining, Over Expression

    The SCN5A DNA promoter region contains conserved Meis1 binding sites. (A) Conservative Meis1 binding sites in the proximal region (2000 nt upstream) of SCN5A DNA promoter computational predicted by JASPAR database (JASPAR 2020 ). (B) ChIP assay was performed by using antibody against Meis1, and IgG as control on the mice ventricular myocardium in sham or MI group. (C-D) Real-time PCR was performed to evaluate the expression of Meis1 mRNA in neonatal mouse cardiomyocytes by knockdown of Meis1 with si-Meis1 or overexpressed of Meis1 by plasmid. At least six independent batches of cells for each group, ** P < 0.01. (E-F) The mRNA and protein levels of sodium channel subunit SCN5A /Na V 1.5 in neonatal mouse cardiomyocytes of NC, si-NC, si-Meis1 and Meis1 group (n = 3–5). Two-tailed student’s t -test was performed. * P < 0.05, ** P < 0.01.
    Figure Legend Snippet: The SCN5A DNA promoter region contains conserved Meis1 binding sites. (A) Conservative Meis1 binding sites in the proximal region (2000 nt upstream) of SCN5A DNA promoter computational predicted by JASPAR database (JASPAR 2020 ). (B) ChIP assay was performed by using antibody against Meis1, and IgG as control on the mice ventricular myocardium in sham or MI group. (C-D) Real-time PCR was performed to evaluate the expression of Meis1 mRNA in neonatal mouse cardiomyocytes by knockdown of Meis1 with si-Meis1 or overexpressed of Meis1 by plasmid. At least six independent batches of cells for each group, ** P < 0.01. (E-F) The mRNA and protein levels of sodium channel subunit SCN5A /Na V 1.5 in neonatal mouse cardiomyocytes of NC, si-NC, si-Meis1 and Meis1 group (n = 3–5). Two-tailed student’s t -test was performed. * P < 0.05, ** P < 0.01.

    Techniques Used: Binding Assay, Real-time Polymerase Chain Reaction, Expressing, Plasmid Preparation, Two Tailed Test

    CDC20 affects the stability of Meis1 in cardiomyocytes. (A) The expression of E3 enzyme related to Meis1 in MI and Sham group mouse hearts (n = 5) analyzed by qRT-PCR assay. * P < 0.05 vs. Sham group. (B) The protein expression of CDC20 in the hearts after 4 weeks of MI (n = 8). Two-tailed student’s t -test was performed. (C) The expression of CDC20 protein was increased in neonatal mouse cardiomyocytes treated with hypoxia for 24 h (n = 4). (D) The variation of Na V 1.5 and Meis1 expression in hypoxic cardiomyocytes with or without knocking down CDC20 by si-CDC20 transfection (n = 4). (E) The variation of Na V 1.5 and Meis1 expression in hypoxic cardiomyocytes after forced expression of CDC20 by CDC20-plasmid transfection (n = 4–5). One-way ANOVA was used to determine statistical significance in these experiments. Error bars represent mean ± SEM of each group. * P < 0.05, ** P < 0.01.
    Figure Legend Snippet: CDC20 affects the stability of Meis1 in cardiomyocytes. (A) The expression of E3 enzyme related to Meis1 in MI and Sham group mouse hearts (n = 5) analyzed by qRT-PCR assay. * P < 0.05 vs. Sham group. (B) The protein expression of CDC20 in the hearts after 4 weeks of MI (n = 8). Two-tailed student’s t -test was performed. (C) The expression of CDC20 protein was increased in neonatal mouse cardiomyocytes treated with hypoxia for 24 h (n = 4). (D) The variation of Na V 1.5 and Meis1 expression in hypoxic cardiomyocytes with or without knocking down CDC20 by si-CDC20 transfection (n = 4). (E) The variation of Na V 1.5 and Meis1 expression in hypoxic cardiomyocytes after forced expression of CDC20 by CDC20-plasmid transfection (n = 4–5). One-way ANOVA was used to determine statistical significance in these experiments. Error bars represent mean ± SEM of each group. * P < 0.05, ** P < 0.01.

    Techniques Used: Expressing, Quantitative RT-PCR, Two Tailed Test, Transfection, Plasmid Preparation

    CDC20 promotes Meis1 ubiquitination and degradation in cardiomyocytes. (A) Flag-tagged Meis1 and His-tagged CDC20 plasmid were overexpressed in HEK293T cells for 48 h, followed by immunoprecipitation with His antibody and western blotting for Flag (n = 3). (B) Immunoprecipitation was performed on the protein extracted from mouse ventricular myocardium by using the antibody against Meis1 and western blotting for CDC20 (n = 3). (C-D) Lysates collected from cardiomyocytes treated with CDC20 plasmid or si-CDC20 and pretreatment with MG132 for 24 h were immunoprecipitated with anti-Meis1. The ubiquitin-conjugated Meis1 was detected by western blotting with anti-ubiquitin (Ub). Input representing the protein expression in whole cell lysates. (E-F) Neonatal mouse cardiomyocytes were treated with CDC20 plasmid or si-CDC20 in the presence or absence of MG132 (5 μM) for 24 h. Representative immunoblotting analyses of Meis1 expression levels for each group (n = 5–6). (G) The expression of Na V 1.5 and Meis1 in hypoxia neonatal mouse cardiomyocytes after knocking down of CDC20 and Meis1 by si-CDC20/si-Meis1 transfection for 24 h (n = 5). One-way ANOVA was used to determine statistical significance in these experiments. Error bars represent mean ± SEM of each group. * P < 0.05, ** P < 0.01.
    Figure Legend Snippet: CDC20 promotes Meis1 ubiquitination and degradation in cardiomyocytes. (A) Flag-tagged Meis1 and His-tagged CDC20 plasmid were overexpressed in HEK293T cells for 48 h, followed by immunoprecipitation with His antibody and western blotting for Flag (n = 3). (B) Immunoprecipitation was performed on the protein extracted from mouse ventricular myocardium by using the antibody against Meis1 and western blotting for CDC20 (n = 3). (C-D) Lysates collected from cardiomyocytes treated with CDC20 plasmid or si-CDC20 and pretreatment with MG132 for 24 h were immunoprecipitated with anti-Meis1. The ubiquitin-conjugated Meis1 was detected by western blotting with anti-ubiquitin (Ub). Input representing the protein expression in whole cell lysates. (E-F) Neonatal mouse cardiomyocytes were treated with CDC20 plasmid or si-CDC20 in the presence or absence of MG132 (5 μM) for 24 h. Representative immunoblotting analyses of Meis1 expression levels for each group (n = 5–6). (G) The expression of Na V 1.5 and Meis1 in hypoxia neonatal mouse cardiomyocytes after knocking down of CDC20 and Meis1 by si-CDC20/si-Meis1 transfection for 24 h (n = 5). One-way ANOVA was used to determine statistical significance in these experiments. Error bars represent mean ± SEM of each group. * P < 0.05, ** P < 0.01.

    Techniques Used: Plasmid Preparation, Immunoprecipitation, Western Blot, Expressing, Transfection