mouse antihuman cd31 pecam 1 antibody  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: A Novel High Content Angiogenesis Assay Reveals That Lacidipine, L-Type Calcium Channel Blocker, Induces In Vitro Vascular Lumen Expansion

doi: 10.3390/ijms23094891

Figure Lengend Snippet: Effect of Lacidipine on the endothelial cell functions. Effect of Lacidipine on cell proliferation in HUVEC cultured as a monolayer ( a ) or in the angiogenesis assay ( b ); n = 6 per concentration, cell count was assessed after 48 h of assay initiation. Data expressed as mean ± SEM, one-way ANOVA followed by Bonferroni’s post hoc test, *** p < 0.001 compared with vehicle. ( c ) Representative fluorescence images following Oris TM migration assay of HUVEC treated with vehicle or Lacidipine (1, 2, and 5 μM) for 22 h. The yellow dotted line represents the cell edge at 0 h after removing the stoppers. Scale bar = 0.5 mm. ( Right panel ): quantification of cell migration. The graph represents the average per cent of gap closure ± S.E.M. from six independent experiments. Unpaired student’s t -test was used. No significant differences between groups were detected. ( d ) The effect of Lacipidine (1, 2, and 5 μM) on tube formation was assessed in a coculture angiogenesis assay where HUVECs were plated on a confluent human dermal fibroblast layer. The medium containing the compound was refreshed on days 3 and 5 following the plating of endothelial cells. Cocultures were stained with an antibody against CD31 and imaged 7 days after endothelial cell plating. The representative fluorescence images of the cocultures treated with Lacidipine (5 μM) and DMSO (vehicle). Inset , “White arrows” indicate the newly formed lumens. ( Left panel ), total tube length was quantified in nine fields for each well ( n = 6 wells per group). Error bars, mean ± SEM, one-way ANOVA followed by Bonferroni’s post hoc test, **** p < 0.001 as compared with vehicle. Scale bar = 100 μm.

Article Snippet: Endothelial tubes were stained using a mouse antihuman CD31 monoclonal antibody (ref. BBA7, R&D systems) at a 1:100 dilution and goat Alexa 568-conjugated anti-mouse IgG secondary antibody (Invitrogen) at a 1:400 dilution.

Techniques: Cell Culture, Angiogenesis Assay, Concentration Assay, Cell Counting, Fluorescence, Migration, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Engineering Vascularized Bone Grafts by Integrating a Biomimetic Periosteum and β-TCP Scaffold

doi: 10.1021/am502056q

Figure Lengend Snippet: HUVECs on an undifferentiated hMSCs sheet formed numerous networks. Networks started at day 3 (A), and elongated to form many lumens at day 5 (B) and day 7 (C). Arrows indicate lumens. Immunofluorescent staining images of CD31 show several networks on the hMSCs sheet at day 7 (D, 10× magnification; E, 20× magnification); 3D-reconstructed confocal images display lumen formation (F). Asterisks show the lumens. Alizarin red staining (G) and vov Kossa staining (H) show the mineralized matrix of osteogenic hMSC cell sheet.

Article Snippet: A 5% goat serum-PBS buffer solution was used to block the cell sheet samples for 1 h at room temperature, and then primary antibody mouse antihuman CD31 (89C2, Cell Signaling Technology, dilution 1:3200) in 1% BSA-PBS was added into the sample, followed by incubation overnight at 4 °C.

Techniques: Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Engineering Vascularized Bone Grafts by Integrating a Biomimetic Periosteum and β-TCP Scaffold

doi: 10.1021/am502056q

Figure Lengend Snippet: Immunohistochemistry staining of human CD31 shows that many antihuman CD31 positive-expressing lumens were seen in HUVEC-UM/OM/β-TCP and OM/HUVEC-UM/β-TCP (black arrows), but there was no expression in plain β-TCP scaffolds and the nonprevascularized OM/UM/β-TCP group (A). A magnified image from HUVEC-UM/OM/β-TCP group at 2 weeks shows that the preformed human blood vessel lumen (brown color, red arrow) carried murine blood cells (semitransparent round ball, blue arrow) (B). The density of the CD31 positive-expressing lumens was higher in HUVEC-UM/OM/β-TCP than that in OM/HUVEC-UM/β-TCP at 2, 4, and 8 weeks (* p < 0.05, n = 4) (C). Immunofluorescent double staining shows the expressions of antihuman CD31 (green) and antimouse CD31 (magenta) in nonprevascularized and prevascularized scaffold group. The overlap or partial overlap of the two colors implies the anastomosis of the preformed human capillaries with the host vasculature (yellow, white arrow) (D).

Article Snippet: A 5% goat serum-PBS buffer solution was used to block the cell sheet samples for 1 h at room temperature, and then primary antibody mouse antihuman CD31 (89C2, Cell Signaling Technology, dilution 1:3200) in 1% BSA-PBS was added into the sample, followed by incubation overnight at 4 °C.

Techniques: Immunohistochemistry, Staining, Expressing, Double Staining