anti pakt ser 473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti pakt ser 473
    Anti Pakt Ser 473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti pakt ser473
    Rabbit Anti Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pakt ser 473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt ser 473
    List of Antibodies for Western Blotting and IHC
    Pakt Ser 473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Hepatocyte Smoothened Activity Controls Susceptibility to Insulin Resistance and Nonalcoholic Fatty Liver Disease"

    Article Title: Hepatocyte Smoothened Activity Controls Susceptibility to Insulin Resistance and Nonalcoholic Fatty Liver Disease

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    doi: 10.1016/j.jcmgh.2022.12.008

    List of Antibodies for Western Blotting and IHC
    Figure Legend Snippet: List of Antibodies for Western Blotting and IHC

    Techniques Used: Western Blot

    anti pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti pakt ser473
    Anti Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt ser473
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    pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt ser473
    HG-induced TGFβ1 activation and signaling are blocked by PI3K/Akt inhibition. HG (48 h)-induced TGFβ1 activation, assessed by ELISA of medium without acid activation, was prevented by PI3K inhibition using either (A) LY294002 ( n = 4, ** p < 0.01, *** p < 0.005) or (B) wortmannin ( n = 4, ** p < 0.01, **** p < 0.0001), as well as Akt inhibition using (C) Akt inhibitor VIII ( n = 4, ** p < 0.01). Smad3 signaling downstream of TGFβ1 was assessed by its C-terminal phosphorylation at <t>Ser473/475</t> (pSmad3). HG (48 h)-induced Smad3 phosphorylation was prevented by (D) LY294002, (E) wortmannin, and (F) Akt inhibitor VIII ( n = 4, * p < 0.05).
    Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Cell surface GRP78 regulates TGFβ1-mediated profibrotic responses via TSP1 in diabetic kidney disease"

    Article Title: Cell surface GRP78 regulates TGFβ1-mediated profibrotic responses via TSP1 in diabetic kidney disease

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2023.1098321

    HG-induced TGFβ1 activation and signaling are blocked by PI3K/Akt inhibition. HG (48 h)-induced TGFβ1 activation, assessed by ELISA of medium without acid activation, was prevented by PI3K inhibition using either (A) LY294002 ( n = 4, ** p < 0.01, *** p < 0.005) or (B) wortmannin ( n = 4, ** p < 0.01, **** p < 0.0001), as well as Akt inhibition using (C) Akt inhibitor VIII ( n = 4, ** p < 0.01). Smad3 signaling downstream of TGFβ1 was assessed by its C-terminal phosphorylation at Ser473/475 (pSmad3). HG (48 h)-induced Smad3 phosphorylation was prevented by (D) LY294002, (E) wortmannin, and (F) Akt inhibitor VIII ( n = 4, * p < 0.05).
    Figure Legend Snippet: HG-induced TGFβ1 activation and signaling are blocked by PI3K/Akt inhibition. HG (48 h)-induced TGFβ1 activation, assessed by ELISA of medium without acid activation, was prevented by PI3K inhibition using either (A) LY294002 ( n = 4, ** p < 0.01, *** p < 0.005) or (B) wortmannin ( n = 4, ** p < 0.01, **** p < 0.0001), as well as Akt inhibition using (C) Akt inhibitor VIII ( n = 4, ** p < 0.01). Smad3 signaling downstream of TGFβ1 was assessed by its C-terminal phosphorylation at Ser473/475 (pSmad3). HG (48 h)-induced Smad3 phosphorylation was prevented by (D) LY294002, (E) wortmannin, and (F) Akt inhibitor VIII ( n = 4, * p < 0.05).

    Techniques Used: Activation Assay, Inhibition, Enzyme-linked Immunosorbent Assay

    Cell surface GRP78 mediates HG-induced TSP1 expression through Akt activation. (A) csGRP78 inhibition using the C38 antibody, but not a control IgG (2 µg for each antibody) prevented HG (24 h)-induced TSP1 transcript upregulation ( n = 6–8, * p < 0.05). HG (48 h)-induced TSP1 promoter activity, assessed using a luciferase reporter construct, was also inhibited by csGRP78 inhibition using either (B) the C38 antibody ( n = 3, ** p < 0.01, *** p < 0.005, **** p < 0.0001) or (C) vaspin ( n = 3, ** p < 0.01, *** p < 0.005, **** p < 0.001). Further, csGRP78 inhibition also prevented HG (48 h)-induced Akt activation assessed by its phosphorylation at Ser473 as well as TSP1 upregulation using (D) C38, but not control IgG, antibody ( n = 5, * p < 0.05, ** p < 0.01), (E) vaspin ( n = 10, * p < 0.05, ** p < 0.01, *** p < 0.005) and (F) siRNA knockdown of MTJ1, the chaperone required for cell surface translocation of GRP78 in HG ( n = 3, * p < 0.05, ** p < 0.01). (G) Antibody neutralization of the known ligand and activator for csGRP78, α2M*, prevented HG (48 h)-induced TSP1 upregulation. Control IgG had no effect (10 µg for each antibody, n = 3, * p < 0.05). (H) An inhibitory peptide (Pep) that prevents the interaction between α2M* and csGRP78 also inhibited TSP1 upregulation by HG (48 h), with scrambled peptide (Scr) having no effect (100 nM, n = 5, * p < 0.05, ** p < 0.01, *** p < 0.005).
    Figure Legend Snippet: Cell surface GRP78 mediates HG-induced TSP1 expression through Akt activation. (A) csGRP78 inhibition using the C38 antibody, but not a control IgG (2 µg for each antibody) prevented HG (24 h)-induced TSP1 transcript upregulation ( n = 6–8, * p < 0.05). HG (48 h)-induced TSP1 promoter activity, assessed using a luciferase reporter construct, was also inhibited by csGRP78 inhibition using either (B) the C38 antibody ( n = 3, ** p < 0.01, *** p < 0.005, **** p < 0.0001) or (C) vaspin ( n = 3, ** p < 0.01, *** p < 0.005, **** p < 0.001). Further, csGRP78 inhibition also prevented HG (48 h)-induced Akt activation assessed by its phosphorylation at Ser473 as well as TSP1 upregulation using (D) C38, but not control IgG, antibody ( n = 5, * p < 0.05, ** p < 0.01), (E) vaspin ( n = 10, * p < 0.05, ** p < 0.01, *** p < 0.005) and (F) siRNA knockdown of MTJ1, the chaperone required for cell surface translocation of GRP78 in HG ( n = 3, * p < 0.05, ** p < 0.01). (G) Antibody neutralization of the known ligand and activator for csGRP78, α2M*, prevented HG (48 h)-induced TSP1 upregulation. Control IgG had no effect (10 µg for each antibody, n = 3, * p < 0.05). (H) An inhibitory peptide (Pep) that prevents the interaction between α2M* and csGRP78 also inhibited TSP1 upregulation by HG (48 h), with scrambled peptide (Scr) having no effect (100 nM, n = 5, * p < 0.05, ** p < 0.01, *** p < 0.005).

    Techniques Used: Expressing, Activation Assay, Inhibition, Activity Assay, Luciferase, Construct, Translocation Assay, Neutralization

    HG-induced TGFβ1 activation requires csGRP78. Assessment of active TGFβ1 in the medium by ELISA showed that csGRP78 inhibition by (A) C38, but not a control IgG antibody ( n = 8, **** p < 0.0001, (B) vaspin ( n = 4, ** p < 0.01, *** p < 0.005) or (C) MTJ1 siRNA knockdown ( n = 4, ** p < 0.01, *** p < 0.005) prevented HG (48 h)-induced TGFβ1 activation. (D–F) MC were co-cultured with mink lung epithelial cells (MLEC) stably transfected with the Smad3-regulated PAI-1 promoter luciferase. The HG (48 h)-induced increase in PAI-1 luciferase activation, reflecting bioactive TGFβ1, was prevented by csGRP78 inhibition with (D) C38, but not control IgG antibody or (E) vaspin, as well as by α2M* inhibition with (F) a neutralizing antibody, but not control IgG (10 µg) or (G) a peptide that prevents csGRP78/α2M* interaction (100 nM, n = 12, * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.0001). In MC, signaling downstream of TGFβ1 was assessed by immunoblotting for Smad3 phosphorylated at Ser473/475. Inhibition of csGRP78 using both (H) vaspin and (I) MTJ1 siRNA knockdown prevented Smad3 activation in HG ( n = 3, * p < 0.05, ** p < 0.01). (J) TSP1 was immunoprecipitated from live cells after HG (48 h) using C38, with IgG used as a control. Immunoblotting shows association with both LAP and csGRP78 in response to HG ( n = 3, **** p < 0.0001).
    Figure Legend Snippet: HG-induced TGFβ1 activation requires csGRP78. Assessment of active TGFβ1 in the medium by ELISA showed that csGRP78 inhibition by (A) C38, but not a control IgG antibody ( n = 8, **** p < 0.0001, (B) vaspin ( n = 4, ** p < 0.01, *** p < 0.005) or (C) MTJ1 siRNA knockdown ( n = 4, ** p < 0.01, *** p < 0.005) prevented HG (48 h)-induced TGFβ1 activation. (D–F) MC were co-cultured with mink lung epithelial cells (MLEC) stably transfected with the Smad3-regulated PAI-1 promoter luciferase. The HG (48 h)-induced increase in PAI-1 luciferase activation, reflecting bioactive TGFβ1, was prevented by csGRP78 inhibition with (D) C38, but not control IgG antibody or (E) vaspin, as well as by α2M* inhibition with (F) a neutralizing antibody, but not control IgG (10 µg) or (G) a peptide that prevents csGRP78/α2M* interaction (100 nM, n = 12, * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.0001). In MC, signaling downstream of TGFβ1 was assessed by immunoblotting for Smad3 phosphorylated at Ser473/475. Inhibition of csGRP78 using both (H) vaspin and (I) MTJ1 siRNA knockdown prevented Smad3 activation in HG ( n = 3, * p < 0.05, ** p < 0.01). (J) TSP1 was immunoprecipitated from live cells after HG (48 h) using C38, with IgG used as a control. Immunoblotting shows association with both LAP and csGRP78 in response to HG ( n = 3, **** p < 0.0001).

    Techniques Used: Activation Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Cell Culture, Stable Transfection, Transfection, Luciferase, Western Blot, Immunoprecipitation

    anti pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti pakt ser473
    Anti Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit monoclonal phospho pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal phospho pakt ser473
    Rabbit Monoclonal Phospho Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti pakt ser473
    Anti Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pakt ser473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pakt ser473
    Membrane stiffness and signaling alteration in the MGO and GAD-treated cultures. ( A ) Confocal images of CTxB labeled cells (F–fibroblast, A–adipocyte), enlarged image highlighted with ICA LUTs to emphasize the cells’ membrane intensity (scale bar = 25 µm). ( B ) Quantification of membrane/cytoplasm CTxB expression ratio at the single-cell level, analyzed in fibroblasts and adipocytes (n = 40–80 cells per group). ( C ) The 8-ANS spectroscopy of the GM culture lysate compared to MGO and GAD-treated cells (n = 6). ( D ) Spectroscopy of treated adipocytes in the presence of Nile red: fluorescence spectrum with blue shift peak (right), and peak value quantification (left, n = 4). ( E ) Western blot analysis of phosphorylated AKT protein (pAKT <t>ser473</t> ) compared to total-AKT in GM, MGO and GAD-treated adipocytes. Results presented as mean ± SD, statistically analyzed by one-way ANOVA. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
    Pakt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Advanced Glycation End Products Effects on Adipocyte Niche Stiffness and Cell Signaling"

    Article Title: Advanced Glycation End Products Effects on Adipocyte Niche Stiffness and Cell Signaling

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms24032261

    Membrane stiffness and signaling alteration in the MGO and GAD-treated cultures. ( A ) Confocal images of CTxB labeled cells (F–fibroblast, A–adipocyte), enlarged image highlighted with ICA LUTs to emphasize the cells’ membrane intensity (scale bar = 25 µm). ( B ) Quantification of membrane/cytoplasm CTxB expression ratio at the single-cell level, analyzed in fibroblasts and adipocytes (n = 40–80 cells per group). ( C ) The 8-ANS spectroscopy of the GM culture lysate compared to MGO and GAD-treated cells (n = 6). ( D ) Spectroscopy of treated adipocytes in the presence of Nile red: fluorescence spectrum with blue shift peak (right), and peak value quantification (left, n = 4). ( E ) Western blot analysis of phosphorylated AKT protein (pAKT ser473 ) compared to total-AKT in GM, MGO and GAD-treated adipocytes. Results presented as mean ± SD, statistically analyzed by one-way ANOVA. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
    Figure Legend Snippet: Membrane stiffness and signaling alteration in the MGO and GAD-treated cultures. ( A ) Confocal images of CTxB labeled cells (F–fibroblast, A–adipocyte), enlarged image highlighted with ICA LUTs to emphasize the cells’ membrane intensity (scale bar = 25 µm). ( B ) Quantification of membrane/cytoplasm CTxB expression ratio at the single-cell level, analyzed in fibroblasts and adipocytes (n = 40–80 cells per group). ( C ) The 8-ANS spectroscopy of the GM culture lysate compared to MGO and GAD-treated cells (n = 6). ( D ) Spectroscopy of treated adipocytes in the presence of Nile red: fluorescence spectrum with blue shift peak (right), and peak value quantification (left, n = 4). ( E ) Western blot analysis of phosphorylated AKT protein (pAKT ser473 ) compared to total-AKT in GM, MGO and GAD-treated adipocytes. Results presented as mean ± SD, statistically analyzed by one-way ANOVA. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

    Techniques Used: Labeling, Expressing, Spectroscopy, Fluorescence, Western Blot

    anti pakt ser 473  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti pakt ser 473
    Anti Pakt Ser 473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    List of Antibodies for Western Blotting and IHC

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Hepatocyte Smoothened Activity Controls Susceptibility to Insulin Resistance and Nonalcoholic Fatty Liver Disease

    doi: 10.1016/j.jcmgh.2022.12.008

    Figure Lengend Snippet: List of Antibodies for Western Blotting and IHC

    Article Snippet: pAkt Ser 473 , Cell Signaling , Cat. #4060 , 1:1000.

    Techniques: Western Blot