rabbit polyclonal antibody against vegfr2  (Cell Signaling Technology Inc)

Journal: Cell Cycle

Article Title: Cancer cell stemness, responses to experimental genotoxic treatments, cytomegalovirus protein expression and DNA replication stress in pediatric medulloblastomas

doi: 10.1080/15384101.2020.1728025

Figure Lengend Snippet: Immunohistochemical data for marker expression in human MB*.

Article Snippet: The primary antibodies used for this immunohistochemical analysis included the following reagents: mouse monoclonal antibody against CD133 (AC133 pure, Miltenyi Biotec, diluted 1:50), mouse monoclonal antibody to CD15 (NCL-CD15, Leica, diluted 1:100), rabbit polyclonal antibody against VEGFR2 (Cell Signaling, diluted 1:250), and rabbit polyclonal antibody to phosphorylated RPA32 (Ser 4/8, NBP1-23017, Novus Biologicals, 1:2000).

Techniques: Immunohistochemical staining, Marker, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: Apatinib, an Inhibitor of Vascular Endothelial Growth Factor Receptor 2, Suppresses Pathologic Ocular Neovascularization in Mice.

doi: 10.1167/iovs.17-21416

Figure Lengend Snippet: FIGURE 1. Apatinib inhibits VEGF-induced phosphorylation of VEGFR2 and ERK in HRMECs. (A) Representative Western blotting images and (B, C) densitometric analysis of phosphorylated VEGFR2 (p-VEGFR2; B) and phosphorylated ERK (p-ERK; C) are illustrated. HRMECs treated with apatinib (1 lM) and untreated cells were stimulated with rhVEGF (100 ng/mL) or PBS for 20 minutes. Cell lysates were subjected to Western blotting using primary IgGs against p-VEGFR2 and p-ERK. The membranes were then stripped and reprobed with IgGs against VEGFR2, ERK, and b-actin. Band intensities of p-VEGFR2 and p-ERK were normalized to the band intensities of VEGFR2 and ERK, respectively (*P < 0.05 versus PBS, #P < 0.05 versus VEGF only, n ¼ 3).

Article Snippet: After quenching endogenous peroxidase activity and blocking with 10% normal goat serum, tissue sections were incubated with primary IgGs against p-VEGFR2 (Origene) or glial fibrillary acidic protein (GFAP; Dako), and then incubated with the appropriate fluorescent secondary IgG.

Techniques: Phospho-proteomics, Western Blot

Journal: Investigative ophthalmology & visual science

Article Title: Apatinib, an Inhibitor of Vascular Endothelial Growth Factor Receptor 2, Suppresses Pathologic Ocular Neovascularization in Mice.

doi: 10.1167/iovs.17-21416

Figure Lengend Snippet: FIGURE 3. Intravitreal injection of apatinib decreases retinal neovascularization in mice with OIR. (A) Schematic diagram of the OIR experiment. On P12, mice received a single intravitreal injection of apatinib (1 lg in 1 lL DMSO) or DMSO (1 lL; contralateral control). Five days later (on P17), the eyes were harvested for further analysis. (B) Representative images of whole-mounted retinas with neovascular tufts highlighted in white, and quantification of retinal neovascularization. Retinal vasculature was visualized by staining with isolectin B4 (red). The extent of retinal neovascularization was calculated by dividing the number of pixels in the neovascular tuft area by the number of pixels in the total retinal area. All data are presented as the mean 6 SEM (**P < 0.01, n ¼ 4 mice per group). Scale bar: 500 lm. (C) Immunofluorescence staining was performed using anti-p-VEGFR2 IgGs (green) and isolectin B4 (red) on cryosections prepared from the eyecups of mice with OIR. The arrowheads indicate the double labeling of p-VEGFR2 and isolectin B4. Sections stained with irrelevant nonspecific IgGs are included as negative controls (NC). The nuclei are shown in blue (DAPI). Representative images were selected from three independent experiments with similar results. Scale bar: 50 lm.

Article Snippet: After quenching endogenous peroxidase activity and blocking with 10% normal goat serum, tissue sections were incubated with primary IgGs against p-VEGFR2 (Origene) or glial fibrillary acidic protein (GFAP; Dako), and then incubated with the appropriate fluorescent secondary IgG.

Techniques: Injection, Control, Staining, Labeling

Journal: Investigative ophthalmology & visual science

Article Title: Apatinib, an Inhibitor of Vascular Endothelial Growth Factor Receptor 2, Suppresses Pathologic Ocular Neovascularization in Mice.

doi: 10.1167/iovs.17-21416

Figure Lengend Snippet: FIGURE 4. Intravitreal injection of apatinib decreases CNV formation in mice with laser-induced CNV. (A) Schematic diagram of the laser-induced CNV experiment. Immediately after laser photocoagulation, mice received a single intravitreal injection of apatinib (1 lg in 1 lL DMSO) or DMSO (1 lL; contralateral control). Two weeks later, the eyes were harvested for further analysis. (B) Representative images of flat-mounted choroids with CNV and quantification of the isolectin B4–positive CNV area. The posterior eyecups composed of RPE, choroid, and sclera were flat-mounted and stained with isolectin B4 (red). The CNV areas were quantified by measuring the fluorescence intensity of images with isolectin B4–positive areas. All data are presented as the mean 6 SEM (**P < 0.01, n¼4 mice per group). (C) Immunofluorescence staining was performed using anti-p-VEGFR2 IgGs (green) and isolectin B4 (red) on cryosections prepared from the eyecups of mice with CNV. The arrowheads indicate the double labeling of p- VEGFR2 and isolectin B4 in CNV lesions (white dashed circles). Sections stained with irrelevant nonspecific IgGs are included as negative controls (NC). The nuclei are shown in blue (DAPI). Representative images were selected from three independent experiments with similar results. Scale bars: 100 lm.

Article Snippet: After quenching endogenous peroxidase activity and blocking with 10% normal goat serum, tissue sections were incubated with primary IgGs against p-VEGFR2 (Origene) or glial fibrillary acidic protein (GFAP; Dako), and then incubated with the appropriate fluorescent secondary IgG.

Techniques: Injection, Control, Staining, Labeling