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 more times with PBS, and the plates were incubated for 1 h at 25°C with 1.0 μg/mL anti-adiponectin antibody (BioVendor, RD181023100) in 1% (w/v) casein-Na blocking solution.

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 more times with PBS, and the plates were incubated for 1 h at 25°C with 1.0 μg/mL anti-adiponectin antibody (BioVendor, RD181023100) in 1% (w/v) casein-Na blocking solution.

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 more times with PBS, and the plates were incubated for 1 h at 25°C with 1.0 μg/mL anti-adiponectin antibody (BioVendor, RD181023100) in 1% (w/v) casein-Na blocking solution.

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 more times with PBS, and the plates were incubated for 1 h at 25°C with 1.0 μg/mL anti-adiponectin antibody (BioVendor, RD181023100) in 1% (w/v) casein-Na blocking solution.

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 more times with PBS, and the plates were incubated for 1 h at 25°C with 1.0 μg/mL anti-adiponectin antibody (BioVendor, RD181023100) in 1% (w/v) casein-Na blocking solution.

Techniques: Comparison

Journal: iScience

Article Title: Adipocyte-released adipomes in Chagas cardiomyopathy: Impact on cardiac metabolic and immune regulation

doi: 10.1016/j.isci.2024.109672

Figure Lengend Snippet: Adiponectin enables selective capture of adipomes from murine white adipose tissue (WAT) (A) Immunoblot analysis of phospho-Perilipin, cleaved Caspase 7 and Annexin V expression in the WAT lysates of infected (20 DPI) and uninfected C57BL/6J mice (n = 4–8 per group). Bar graph values were derived from densitometry analysis and by normalizing target protein expression to GDI. Error bars indicate the standard error of the mean (∗ p < 0.05). (B) Immunoblot analysis of adiponectin in WAT-derived large-EVs (L-EV) and small-EVs (S-EV). Black arrow points to the 25 kDa marker band on the pre-stained protein ladder. Adiponectin band appears at 26/30 kDa. Lanes: C, control WAT lysate; L Ev, large-EV; and S Ev, small-EV. (C) Surface immunofluorescence staining of 3T3-L1 adipocytes show the co-localization of Adiponectin (green) with Annexin V (red) (top panel) and FABP4 (red) with Annexin V (green) (bottom panel) on budding apoptotic bodies. Scale bar, 50 μm. (D) Immunoblot analysis of adiponectin in WAT-derived large-adipomes and small-adipomes. Black arrow points to the 25 kDa marker band on pre-stained protein ladder. Adiponectin band appears at 26/30 kDa. Lanes: C, control WAT lysate; L, large-adipomes; and S, small-adipomes. (E) Immunofluorescence assay (IFA) of bead-bound adipomes stained with Adiponectin-AF488 (top panel) and FABP4-AF488 (bottom panel) imaged on the FITC channel along with their corresponding bright field (BF) images. Scale bar, 5 μm. (F) WAT-derived adipome size distribution and absolute concentration as determined by Spectradyne nCS1 with a C-2000 microfluidic cartridge. Inset: L (blue), large-adipomes; S (green), small-adipomes. Gate: G1, particle concentration at size range of 250–400 nm diameter showing more small-adipomes than large-adipomes; and G2, particle concentration at size range of 700–1100 nm diameter showing mostly large-adipome distribution. (G) Transmission Electron Microscopy (TEM) images of small-adipomes (left, scale bar 80 nm) and large-adipomes (right, scale bar 100 nm) with a direct magnification of 20,000×. Black arrow indicates the budding scars on L-adipomes.

Article Snippet: Rabbit Polyclonal Anti-Adiponectin , BioVision , Cat#5901-250; RRID: AB_442498.

Techniques: Western Blot, Expressing, Infection, Derivative Assay, Marker, Staining, Immunofluorescence, Concentration Assay, Transmission Assay, Electron Microscopy

Journal: iScience

Article Title: Adipocyte-released adipomes in Chagas cardiomyopathy: Impact on cardiac metabolic and immune regulation

doi: 10.1016/j.isci.2024.109672

Figure Lengend Snippet: Intracardiac P-ILA treatment increases cardiac adiponectin (APN) and Atrial natriuretic peptide (ANP) in murine CD model (A) Immunoblot analysis of adiponectin and β1-AR expression in heart lysates of uninfected and post-acute T. cruzi -infected mice treated with intracardiac injections of either P-ILA or vehicle. Bar graph values were derived from densitometry analysis and by normalizing target protein expression to GDI. Lanes: 1, uninfected vehicle-injected; 2, uninfected P-ILA-injected; 3, T. cruzi -infected vehicle-injected; and 4, T. cruzi -infected P-ILA-injected murine heart tissue lysate. The “&” symbol indicates significance calculated between uninfected and T. cruzi -infected P-ILA injections. (B) Representative immunohistochemistry (IHC) images of vehicle- and P-ILA-injected heart tissue from uninfected and T. cruzi -infected mice showing ANP immunostaining visualized with DAB and counterstained with hematoxylin. Dot plot shows staining intensity in O.D. (mean gray value) obtained by color deconvolution analysis.Data are represented as mean ± SEM. (∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001).

Article Snippet: Rabbit Polyclonal Anti-Adiponectin , BioVision , Cat#5901-250; RRID: AB_442498.

Techniques: Western Blot, Expressing, Infection, Derivative Assay, Injection, Immunohistochemistry, Immunostaining, Staining

Journal: iScience

Article Title: Adipocyte-released adipomes in Chagas cardiomyopathy: Impact on cardiac metabolic and immune regulation

doi: 10.1016/j.isci.2024.109672

Figure Lengend Snippet:

Article Snippet: Rabbit Polyclonal Anti-Adiponectin , BioVision , Cat#5901-250; RRID: AB_442498.

Techniques: Infection, Recombinant, Electron Microscopy, SYBR Green Assay, Lysis, Protease Inhibitor, Plasmid Preparation, XF Assay, Isolation, Quantitation Assay, Bicinchoninic Acid Protein Assay, Software

Journal: Bone & Joint Research

Article Title: Adiponectin inhibits fibrosis of the palmar aponeurosis in Dupuytren’s contracture in male patients

doi: 10.1302/2046-3758.128.BJR-2022-0449.R1

Figure Lengend Snippet: Primer sequences for quantitative real-time polymerase chain reaction.

Article Snippet: The sections were then incubated overnight at 4°C with a rabbit anti-human adiponectin polyclonal antibody (21613-1-AP, Proteintech, USA; dilution 1:200) and a rabbit anti-human leptin polyclonal antibody (ab16227, Abcam, USA; dilution 1:100).

Techniques:

Journal: Bone & Joint Research

Article Title: Adiponectin inhibits fibrosis of the palmar aponeurosis in Dupuytren’s contracture in male patients

doi: 10.1302/2046-3758.128.BJR-2022-0449.R1

Figure Lengend Snippet: a) Adiponectin gene expression was significantly decreased in the Dupuytren’s contracture group compared to that in the carpal tunnel syndrome (CTS) group (p < 0.001) (n = 8). Gene expression was normalized against the CTS. All results are expressed as means and standard error. b) Fluorescence immunohistochemical staining showing decreased protein expression in the Dupuytren’s contracture group compared to the CTS group (20×). H&E, haematoxylin and eosin.

Article Snippet: The sections were then incubated overnight at 4°C with a rabbit anti-human adiponectin polyclonal antibody (21613-1-AP, Proteintech, USA; dilution 1:200) and a rabbit anti-human leptin polyclonal antibody (ab16227, Abcam, USA; dilution 1:100).

Techniques: Gene Expression, Fluorescence, Immunohistochemical staining, Staining, Expressing