recombinant adiponectin  (BioVendor Instruments)

Journal: World Journal of Hepatology

Article Title: Transforming growth factor beta reduces proprotein convertase subtilisin/kexin type 9 in the supernatant of hepatic stellate cells

doi: 10.4254/wjh.v18.i1.113896

Figure Lengend Snippet: Regulation of proprotein convertase subtilisin/kexin type 9 levels in LX-2 cells by cytokines, adipokines, and nuclear receptor agonists. A and B: Proprotein convertase subtilisin/kexin type 9 (PCSK9) levels in LX-2 cells cultivated with tumor necrosis factor and interleukin-6. PCSK9 levels in the LX-2 cell supernatant after 24 hours of cultivation with 0 ng/mL, 0.2 ng/mL, 2 ng/mL, or 4 ng/mL tumor necrosis factor, 4 independent experiments (A); PCSK9 levels in the LX-2 cell supernatant after 24 hours of cultivation with 0 ng/mL, 5 ng/mL, 10 ng/mL, or 20 ng/mL interleukin-6, 4 independent experiments (B); C-E: PCSK9 levels during LX-2 cell cultivation with adipokines and chemerin isoform overexpression. PCSK9 levels in the LX-2 cell culture supernatant after 24 hours of cultivation with 0 μg/mL or 10 μg/mL adiponectin, 6 independent experiments (C); PCSK9 levels in the LX-2 cell culture supernatant after 24 hours of cultivation with 0 ng/mL, 4 ng/mL, 10 ng/mL, or 20 ng/mL leptin, 4 independent experiments (D); PCSK9 in the supernatant of LX-2 cells cultivated for 24 hours with overexpressed chemerin-155, chemerin-156, or chemerin-157 and in control transfected cells, 4 independent experiments (E); F-H: PCSK9 levels during LX-2 cell cultivation with nuclear receptor agonists. PCSK9 levels in the LX-2 cell culture supernatant after 24 hours of cultivation with 0 μmol/L, 1 μmol/L, and 2 μmol/L GW4064, 5 independent experiments (F); PCSK9 levels in the LX-2 cell culture supernatant after 24 hours of cultivation with 0 μmol/L, 5 μmol/L, and 10 μmol/L T0901317, 3 independent experiments (G); PCSK9 in the supernatant of LX-2 cells cultivated for 24 hours with 0 μmol/L, 7.5 μmol/L, and 15 μmol/L rosiglitazone, 3 independent experiments (H). PCSK9: Proprotein convertase subtilisin/kexin type 9; TNF: Tumor necrosis factor; IL: Interleukin; Chem: Chemerin.

Article Snippet: Recombinant human adiponectin and leptin were obtained from R&D Systems (Wiesbaden-Nordenstadt, Germany).

Techniques: Over Expression, Cell Culture, Control, Transfection

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