anti-integrin αiib  (Santa Cruz Biotechnology)

Journal: Cells

Article Title: ADAMTS13 Improves Endothelial Function and Reduces Inflammation in Diabetic Retinopathy

doi: 10.3390/cells14020085

Figure Lengend Snippet: Hemostasis biomarkers in vitreous fluid of diabetes patients. Detection of thrombotic markers in vitreous fluid of patients with proliferative diabetic retinopathy (PDR). Determination of the von Willebrand factor (VWF) ( A ), the platelet marker CD41 ( B ), and ADAMTS13 ( C ) levels in vitreous fluid samples. A total of 15 µL of vitreous fluid samples from 12 patients with PDR and from 12 nondiabetic patients with rhegmatogenous retinal detachment (RD) was subjected to gel electrophoresis and the presence of VWF, CD41, and ADAMTS13 (5C11 monoclonal antibody) was illustrated by representative western blots and the levels of the antigens compared between the RD and PDR cohorts. Results are expressed as medians (interquartile range). (* p < 0.05; Mann-Whitney test).

Article Snippet: The immunodetection of specific molecules was conducted with the following reagents and conditions: VWF with a mouse monoclonal anti-VWF antibody (1:1000, sc-365712, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), CD41 with a mouse monoclonal anti-CD41 antibody (1:1000, sc-365938, Santa Cruz Biotechnology Inc.), β-catenin with a goat polyclonal anti-ß-catenin antibody (1:1000, AF1329, R&D system, Minneapolis, MN, USA), ADAMTS13 with a rabbit monoclonal anti-ADAMTS13 antibody (1:1000, NBP3-16038, Novus Biologicals, Littleton, CO, USA) and with the three mouse monoclonal antibodies 3H9, 5C11, and 12H6, as described above, HMGB1 with a rabbit polyclonal anti-high mobility group box-1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam, Cambridge, UK), ERK1/2 with a rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), ICAM-1 with a mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and VCAM-1 with a mouse monoclonal anti-vascular cell adhesion-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Marker, Nucleic Acid Electrophoresis, Western Blot, MANN-WHITNEY

Journal: Cells

Article Title: ADAMTS13 Improves Endothelial Function and Reduces Inflammation in Diabetic Retinopathy

doi: 10.3390/cells14020085

Figure Lengend Snippet: ADAMTS13 expression levels in diabetic rat retinas and effects of exogenously administered ADAMTS13. ADAMTS13 expression levels in the retinal lysates of diabetic rats ( D ) ( n = 12) and nondiabetic control animals ( n = 12) were determined by western blot analysis. After the measurement of the intensities of ADAMTS13 proteoform bands, the immunoblots were stripped and reprobed to evaluate ß-tubulin intensities in all sample panels ( A ). Results are expressed as means ± standard deviation of the ratios between ADAMTS13 and ß-tubulin (* p < 0.05; independent t-test). The effects of intravitreal ADAMTS13 injection on vascular permeability and markers of hemostasis and inflammation in rat retinas after streptozotocin-induced diabetes were evaluated by quantifications of the BRB breakdown by detection of FITC dextran seeped into the retina after the systemic injection ( B ). Retinal protein expression levels of the von Willebrand factor (VWF) ( C ), the platelet marker CD41 ( D ), vascular endothelial (VE)-cadherin ( E ), and ß-catenin ( F ) were determined by immunoblot analysis. Statistical comparisons (mean ± standard deviation of 8–10 rats) were performed as described in . * p < 0.05 compared with values obtained from nondiabetic controls. # p < 0.05 compared with values obtained from diabetic rats.

Article Snippet: The immunodetection of specific molecules was conducted with the following reagents and conditions: VWF with a mouse monoclonal anti-VWF antibody (1:1000, sc-365712, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), CD41 with a mouse monoclonal anti-CD41 antibody (1:1000, sc-365938, Santa Cruz Biotechnology Inc.), β-catenin with a goat polyclonal anti-ß-catenin antibody (1:1000, AF1329, R&D system, Minneapolis, MN, USA), ADAMTS13 with a rabbit monoclonal anti-ADAMTS13 antibody (1:1000, NBP3-16038, Novus Biologicals, Littleton, CO, USA) and with the three mouse monoclonal antibodies 3H9, 5C11, and 12H6, as described above, HMGB1 with a rabbit polyclonal anti-high mobility group box-1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam, Cambridge, UK), ERK1/2 with a rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), ICAM-1 with a mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and VCAM-1 with a mouse monoclonal anti-vascular cell adhesion-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Control, Western Blot, Standard Deviation, Injection, Permeability, Marker