polyclonal goat anti cd31 primary antibody  (R&D Systems)

Journal: Bioactive materials

Article Title: 3D-printed scaffolds with 2D hetero-nanostructures and immunomodulatory cytokines provide pro-healing microenvironment for enhanced bone regeneration.

doi: 10.1016/j.bioactmat.2023.03.021

Figure Lengend Snippet: Fig. 5. The in vitro vascularization and osteogenesis. a) Schematic demonstration of the co-culture of macrophages in IL-4 released medium from the immuno-functionalized 3D scaffolds and the subse quent induction of vascularization for HUVEC cells or osteogenesis for rBMSCs using the supernatant from the macrophage co-culture. Relative mRNA expres sions of b) ANG1, c) bFGF2, d) eNOS3, e) VEGFA, and f) VEGFC vascular marker genes in HUVECs. g) Fluorescence intensities of the CD31 vascular marker and h) confocal images of HUVECs stained immuno fluorescent with CD31 (pink), F-actin (red), vinculin (green), and nuclei (blue: DAPI) after co-culture with macrophage supernatant. Relative mRNA expressions of i) OPN, j) OCN, k) Runx2, and l) OSX osteogenic marker genes in rBMSCs. m) Relative ALP activities, n) fluorescence intensities of Runx2, and o) confocal images of rBMSCs stained with immunofluorescent F- actin (red), Runx2 (green), and nuclei (blue: DAPI) after co-culture with microphage supernatant. (*: p < 0.05).

Article Snippet: The intracellular expression of vascular marker protein CD31 was visualized by immunofluorescence staining using goat anti-CD31 polyclonal primary antibody (Novus Biologicals, CO) and bovine anti-goat IgG CFTM633 secondary antibody (Sigma Aldrich).

Techniques: In Vitro, Co-Culture Assay, Marker, Fluorescence, Staining

Journal: Bioactive materials

Article Title: 3D-printed scaffolds with 2D hetero-nanostructures and immunomodulatory cytokines provide pro-healing microenvironment for enhanced bone regeneration.

doi: 10.1016/j.bioactmat.2023.03.021

Figure Lengend Snippet: Fig. 7. Immunohistological analysis and in vivo neo vascularization and osteogenesis in rat calvarial defect model. a) H & E staining, b) toluidine blue staining, and c) Masson’s trichrome staining of the empty bone defects and defects implanted with 3D- printed scaffolds. d) Immunohistochemical staining of ALP and CD31 proteins in the empty bone defects and defects implanted with 3D-printed scaffolds. Quantitative analysis of in vivo e) ALP and f) CD31 fluorescence intensity in the defect sites. (*: p < 0.05).

Article Snippet: The intracellular expression of vascular marker protein CD31 was visualized by immunofluorescence staining using goat anti-CD31 polyclonal primary antibody (Novus Biologicals, CO) and bovine anti-goat IgG CFTM633 secondary antibody (Sigma Aldrich).

Techniques: In Vivo, Staining, Immunohistochemical staining, Fluorescence

Journal: ACS biomaterials science & engineering

Article Title: Bioorthogonal "Click Chemistry" Bone Cement with Bioinspired Natural Mimicking Microstructures for Bone Repair.

doi: 10.1021/acsbiomaterials.2c01482

Figure Lengend Snippet: Figure 6. Angiogenesis. (a) Proliferation of HUVECs on the porous click-ON cements. (b) HUVEC morphology and distribution in the porous click cements with varied pore sizes. (c) mRNA expression levels of vascular markers in HUVECs growing on the cements. (d) Immunofluorescence staining of the vascular marker CD31 protein in HUVECs growing on the cements. (e) Schematic demonstration of potential support for vascular differentiation of stem cells on the porous click-ON cement and the mRNA expression level of vascular markers of (f) CD31, (g) α-SMA, (h) VEGFA, and (i) VEGFR2 in MSCs growing on the cements. (j) Immunofluorescence imaging of vascular marker CD31 protein expression in MSCs on the three types of click cements.

Article Snippet: To determine the local neovascularization in the rat cranial defect site, immunofluorescence co-staining was performed by incubating tissue sections with rabbit monoclonal anti-ALP primary antibodies (Novus Biologicals) and goat polyclonal anti-rat CD31 primary antibodies (Novus Biologicals).

Techniques: Expressing, Immunofluorescence, Staining, Marker, Imaging

Journal: ACS biomaterials science & engineering

Article Title: Bioorthogonal "Click Chemistry" Bone Cement with Bioinspired Natural Mimicking Microstructures for Bone Repair.

doi: 10.1021/acsbiomaterials.2c01482

Figure Lengend Snippet: Figure 8. In vivo osteogenesis and neovascularization. (a) In vivo osteogenic marker expression and (b) in vivo ALP activities in rat calvarial defect sites. Immunofluorescence intensity quantification of (c) ALP osteogenic maker and (d) CD31 vascular marker in tissue slices from the rat cranial defects with the PO-click-ON cement and the empty control. Immunohistochemistry (IHC)-stained images of ALP (green)/nuclei (blue) and CD31 (red)/nuclei (blue) in tissue slices from (e, f) the empty rat cranial defects and (g, h) the defects with the PO-click-ON cement. (i) Schematic demonstration of limited bone repair in the empty bone defect and the robust enhancement of bone repair by the PO-click-ON cement taking advantage of favorable cell recruitment, osteoinductivity, osteoconductivity, and neovascularization. Asterisk (*): statistically different (p < 0.05).

Article Snippet: To determine the local neovascularization in the rat cranial defect site, immunofluorescence co-staining was performed by incubating tissue sections with rabbit monoclonal anti-ALP primary antibodies (Novus Biologicals) and goat polyclonal anti-rat CD31 primary antibodies (Novus Biologicals).

Techniques: In Vivo, Marker, Expressing, Immunofluorescence, Control, Immunohistochemistry, Staining

Journal: Science Advances

Article Title: A single intravenous injection of cyclosporin A–loaded lipid nanocapsules prevents retinopathy of prematurity

doi: 10.1126/sciadv.abo6638

Figure Lengend Snippet: ( A ) Nanocapsule accumulation after intravenous administration and 1 hour of blood circulation in % ID per gram of tissue in healthy mice. ( B ) Absolute target tissue distribution in % ID of RGD-LNCs and LNCs in the retina. ( C ) TEM images showing dual RGD-LNCs in the posterior eye 1 hour after intravenous administration. Top: RGD-LNCs are associated with endothelial cells. Middle: RGD-LNCs adjacent to and migrating in the Bruch’s membrane. Bottom: RGD-LNCs internalized in RPE cells. Arrows indicate electron-dense superparamagnetic iron oxide nanoparticles (SPIONs) containing dual RGD-LNCs. From left to right: Squared out regions show progressive magnification. Scale bars, 250 nm. Representative images of at least n = 5 biologically independent samples. ( D ) Sagittal section of eyes stained for endothelial cells (CD31; green) and cell nuclei (DAPI; blue) showing RPE-specific accumulation of RGD-LNCs 1 hour after intravenous administration. Nanocapsule accumulation in the RPE is indicated with white arrows. Lilac: Nanocapsule-associated fluorescence. Squared out region shows magnification. Scale bars, 20 μm. ( E and F ) Quantitative analysis of nanoparticle fluorescence 1 hour after administration of LNCs or RGD-LNCs (E) and of RGD-LNCs at different time points (F). Results are presented as means ± SD of n = 6 (A and B) mice per treatment group, n = 5 (E), or at least n = 3 (F). Levels of statistical significance are indicated as * P ≤ 0.05, ** P ≤ 0.01, and **** P ≤ 0.0001. P values were determined by two-way ANOVA (A), unpaired t test (B), and one-way ANOVA (E and F).

Article Snippet: Sections were then washed with phosphate buffer (3×, 5 min each) and incubated overnight with primary polyclonal goat anti-CD31 antibody (R&D Systems, Minneapolis, USA) (1:100 in 1:10 blockage buffer) at 4°C.

Techniques: Membrane, Staining, Fluorescence

Journal: BMC Cancer

Article Title: VEGFR2 heterogeneity and response to anti-angiogenic low dose metronomic cyclophosphamide treatment

doi: 10.1186/1471-2407-10-683

Figure Lengend Snippet: Microvessel density (MVD) as CD31 positive profiles in SW480 and WM239 subcutaneous xenografts; Mean (SEM) are plotted . There were significant reductions in MVD in both SW480 (A) and WM239 (B) xenografts between CTX and control groups (*p < 0.05). Western blotting of tumor lysates for TSP-1 revealed significant increases in TSP-1 levels in CTX treated SW480 tumors compared to control (C; *p < 0.05), but significant decreases in CTX treated WM239 tumors compared to control (D; *p < 0.05).

Article Snippet: Antigen detection was performed sequentially by first using goat polyclonal anti-CD31 primary antibody (1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) then donkey anti-goat FITC-conjugated secondary antibody (1:200; Santa Cruz Biotechnology) for 30 minutes.

Techniques: Control, Western Blot

Journal: BMC Cancer

Article Title: VEGFR2 heterogeneity and response to anti-angiogenic low dose metronomic cyclophosphamide treatment

doi: 10.1186/1471-2407-10-683

Figure Lengend Snippet: Effect of low dose metronomic cyclophosphamide therapy on vascular mural cell recruitment . A) Dual immunofluorescent staining for vascular alpha smooth muscle actin (α-SMA; red) and CD31 (green) showing blood vessels positive (chevrons) and negative (arrows) for this mural cell marker. Scale bar = 50 μm. B) CTX treated SW480 xenograft tumors had significantly increased proportion of α-SMA positive blood vessels compared to control (*p < 0.05). No significant difference was observed between CTX and control WM239 tumors. The ratio between the mural cell marker protein desmin and pan endothelial marker CD31 was assessed by western blotting of whole tumor lysates (C) LDM CTX treatment resulted in increased desmin content realtive to CD31 for SW480 but not WM239 xenografts.

Article Snippet: Antigen detection was performed sequentially by first using goat polyclonal anti-CD31 primary antibody (1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) then donkey anti-goat FITC-conjugated secondary antibody (1:200; Santa Cruz Biotechnology) for 30 minutes.

Techniques: Staining, Marker, Control, Western Blot