recombinant adiponectin  (BioVendor Instruments)

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: (A) Cumulative concentration response curve of methylcholine-induced vasodilation in pre-constricted (with U46619) mesenteric arteries incubated overnight without stimulus (white), 50 µg/mL of oxLDL (red), 30 µg/mL of AN (green), or 30 µg/mL of AN plus 50 µg/mL of oxLDL (blue). (B) Summary graph of the maximum response (Emax) obtained from concentrations curves shown in (A). Vessels were obtained from rats ( n = 6-13). (C) Cumulative concentration response curve of methylcholine-induced vasodilation in pre-constricted (with U46619) mesenteric arteries incubated overnight without stimulus (white) or with 50 µg/mL of oxLDL (red), 30 µg/mL of AN plus 50 µg/mL of oxLDL (blue), 30 µg/mL of AN plus 10 µg/mL of TS20 (purple), or 10 µg/mL of the IgG isotype (black). (D) Summary graph of the maximum response (Emax) obtained from concentrations curves shown in C. Vessels from n = 6-7 rats. Values are shown as the mean±SEM. ＊ p ＜0.05; ＊＊ p ＜0.01; ＊＊＊ p ＜0.001. ns, non-significant.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Concentration Assay, Incubation

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: (A) Schematic representation of recombinant human T-cadherin proteins fused with Myc-His-tag. (B) Coomassie brilliant blue (CBB) staining and western blot analysis of purified T-cadherin proteins. (C) Binding of recombinant AN to immobilized T-cadherin in ELISA was determined by anti-AN antibody. (D) Binding of recombinant AN−oxLDL complex to immobilized LOX-1 in ELISA was determined by anti-AN antibody. s.p., signal peptide; EC, extracellular cadherin.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Staining, Western Blot, Purification, Binding Assay

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: Correlation between total-AN and high-molecular weight (HMW)-AN (left), total-AN and T-cadherin bindable adiponectin (Tcad-AN) (middle), HMW-AN and Tcad-AN (right).

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Enzyme-linked Immunosorbent Assay, High Molecular Weight

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: (A) Serum levels of AN, T-cadherin bindable adiponectin (Tcad-AN), oxidized LDL (oxLDL) and LOX-1-ligand containing apoB (LAB) in patients on hemodialysis (HD; n = 35) and control subjects ( n = 35). The error bars represent the mean±SD of the values obtained. (B) Comparison of the proportion of MAC-high between HD and controls. MAC-high and -low groups were divided by the median of the MAC values of 70 subjects.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Comparison, Control

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: The error bars represent mean±SD.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Comparison

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: Adiponectin inhibits collagen and alpha-smooth muscle actin gene expression . Confluent control ( A-C ) or scleroderma ( D ) dermal fibroblasts were incubated with adiponectin (5 ug/ml) for 24 to 36 hours. A . Total RNA was subjected to real-time qPCR. The results represent the means ± SEM of triplicate determinations. * P <0.05, ** P <0.01. B, D . Whole cell lysates were analyzed by Western blot. Representative autoradiographs. C . Fibroblasts were immunostained with antibodies to Type I collagen (red) or α-SMA (green), or stained with DAPI (blue), and examined by confocal microscopy. Representative immunofluorescence photomicrographs. Original magnification × 60. α-SMA, α-smooth muscle actin; DAPI, 4',6-diamidino-2-phenylindole; SEM, standard error of the mean.

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Expressing, Incubation, Western Blot, Staining, Confocal Microscopy, Immunofluorescence

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: Adiponectin attenuates TGF-β-induced profibrotic responses . A . Confluent dermal fibroblasts were incubated with or without TGF-β (2 ng/ml) in the presence of the indicated concentration of adiponectin for 48 hours. Total RNA was subjected to real-time qPCR. The results represent the means ± SEM of triplicate determinations. B . Fibroblasts were preincubated with adiponectin-specific siRNA or control siRNA. SEM, standard error of the mean; TGF-β, transforming growth factor β

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Incubation, Concentration Assay

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: AMP kinase mediates suppression of profibrotic responses by adiponectin . Confluent dermal fibroblasts in two-dimensional monolayer cultures were incubated with adiponectin (5 ug/ml) in media with TGF-β (2 ng/ml) in the presence or absence of Compound C (10 uM) for 48 hours. Total RNA was harvested and subjected to real-time qPCR. The results represent the means ± SEM of triplicate determinations. SEM, standard error of the mean; TGF-β, transforming growth factor-β.

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Incubation

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: Adiponectin mediates the anti-fibrotic effects of PPAR - γ ligands . Dermal fibroblasts from adiponectin-null mice and wild-type littermates in parallel were incubated with indicated concentrations of PGJ 2 , followed by TGF-β (2 ng/ml) for 24 hours. Total RNA was subjected to real-time qPCR. The results represent the means ± SEM of triplicate determinations. PGJ 2 , prostaglandin J 2 ; SEM, standard error of the mean; TGF-β, transforming growth factor-β.

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Incubation

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: Adiponectin attenuates LPS-induced profibrotic responses . Confluent dermal fibroblasts were incubated with LPS (1 ug/ml) in the absence or presence of adiponectin (10 ug/ml) for 24 hours. Total RNA was subjected to real-time qPCR. The results represent the means ± SEM of triplicate determinations. LPS, lipopolysaccharide; SEM, standard error of the mean.

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Incubation

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: Adiponectin receptor expression in scleroderma. A . Total RNA was isolated from confluent fibroblasts explanted from scleroderma patient ( n = 4) and healthy controls ( n = 4), and subjected to real-time qPCR. The results represent the mean ± SEM of triplicate determinations. B . AdipoR1/2 mRNA expression was interrogated in publicly available genome-wide expression scleroderma skin biopsy microarray datasets (GEO accession number: GSE9285) . Fold-change in mRNA levels was normalized with average expression levels of the samples in the entire cohort. Box plots indicate the range of lower and upper quartiles. * P <0.05. SEM, standard error of the mean.

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Expressing, Isolation, Genome Wide, Microarray

Journal: Arthritis Research & Therapy

Article Title: The adipokine adiponectin has potent anti-fibrotic effects mediated via adenosine monophosphate-activated protein kinase: novel target for fibrosis therapy

doi: 10.1186/ar4070

Figure Lengend Snippet: Adiponectin suppresses fibrotic responses . Ligands of PPAR-γ induce adiponectin which triggers autocrine or paracrine activation of AMP kinase. AMP kinase in turn blocks TGF-β/Smad signaling and consequent stimulation of profibrotic gene expression. PPAR-γ, peroxisome proliferator activated receptor gamma; TGF-β, transforming growth factor-β.

Article Snippet: When fibroblasts reached confluence, growth media with 10% FBS or serum-free media supplemented with 0.1% BSA were added to the cultures for 24 hours prior to TGF-β2 (Peprotech, Rocky Hill, NJ, USA), or full-length adiponectin (Bio Vendor, Karasek, Czech Republic).

Techniques: Activation Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Association between clinical characteristics and serum adiponectin levels in NPC patients.

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: The association of serum adiponectin levels with survival in NPC patients. Kaplan–Meier curves were used to illustrate several patients. ( A ) Overall survival and ( B ) metastasis-free survival were analyzed according to serum adiponectin levels based on a cutoff of 8.00 μg/mL in NPC patients ( n = 106). p -values were determined by the log-rank test.

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Univariable and multivariable Cox regression analyses of potential factors for the metastasis-free survival in NPC patients a .

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Adiponectin suppresses NPC cells invasion and migration. ( A ) Wound healing assay of S18 and 5-8F cells pretreated with adiponectin (0, 10, 20, or 40 μg/mL) for 24 h. Scale bar: 200 μm. Graphs show the relative wound closure. ( B ) Invasion of S18 and 5-8F cells were evaluated by Transwell invasion assay covered with Matrilgel after treatment with various dosing of adiponectin for 24 h. Scale bar: 200 μm. Graphs show the relative number of invasion cells. ( C ) Migration of S18 and 5-8F cells were evaluated by Transwell migration assay after treatment with various doses of adiponectin for 24 h. Scale bar: 200 μm. Graphs show the relative number of migration cells. ( D ) Western blot analysis of E-cadherin, N-cadherin, Slug, Snail, Vimentin, Claudin-1, MMP-2, and MMP-9 in S18 and 5-8F cells after treatment with 40 μg/mL adiponectin for 48 h. Results are presented as mean ± SD of three independent experiments performed in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.0001.

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Migration, Wound Healing Assay, Transwell Invasion Assay, Transwell Migration Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Adiponectin blocks leptin, IL-6, or TNF-α-induced NPC cell invasion and migration. Transwell migration ( A ) and invasion ( B ) assays in S18 and 5-8F cells pretreated with 40 μg/mL adiponectin for 2 h before treatment with 50 ng/mL leptin. Scale bar: 200 μm. Graphs show the relative number of cells. Transwell migration assay in S18 and 5-8F cells pretreated with 40 μg/mL adiponectin for 2 h before treatment with 50 ng/mL TNF-α ( C ) or 50 ng/mL IL-6 ( D ). Scale bar: 200 μm. Graphs show the relative number of migration cells. ( E – G ) Western blot analysis of E-cadherin, N-cadherin, MMP-2, and Vimentin in 5-8F cells pretreated with 40 μg/mL adiponectin in the presence or absence of leptin (50 ng/mL), TNF-α (50 ng/mL), or IL-6 (50 ng/mL) for 48 h. Results are presented as mean ± SD of three independent experiments performed in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.0001.

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Migration, Transwell Migration Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Adiponectin inhibits NF-κB and STAT3 activation in NPC cells. Luciferase reporter assay of NF-κB ( A ) and STAT3 ( B ) in S18 and 5-8F cells after treatment with 40 μg/mL adiponectin for 3 h. ( C ) Western blot analysis of p-IκBα, cyto-p65, and nuc-p65 in S18 and 5-8F cells after treatment with various doses of adiponectinfor 6 h. ( D ) Western blot analysis of p-STAT3, cyto-STAT3, and nuc-STAT3 in S18 and 5-8F cells after treatment with various doses of adiponectin for 6 h. Results are presented as mean ± SD of three independent experiments performed in triplicate. ** p < 0.01, *** p < 0.001.

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Activation Assay, Luciferase, Reporter Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: AdipoR1 mediates the metastasis inhibitory effect of adiponectin in NPC cells. ( A ) The mRNA expression of AdipoR1 and AdipoR2 was measured with qRT-PCR in S18 and 5-8F cells after transfected with AdipoR1 or AdipoR2 siRNA. ( B ) Transwell migration assay in S18 and 5-8F cells treated with 40 μg/mL adiponectin for 24 h after transfected with AdipoR1 or AdipoR2 siRNA. Scale bar: 200 μm. Graphs show the relative number of migration cells. ( C ) Western blot analysis of E-cadherin, N-cadherin, MMP-9, and MMP-2 in S18 and 5-8F cells treated with 40 μg/mL adiponectin for 48 h after transfected with AdipoR1 siRNA. Results are presented as mean ± SD of three independent experiments performed in triplicate. *** p < 0.001, # p < 0.0001.

Article Snippet: Recombination human adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Transwell Migration Assay, Migration, Western Blot