primary antibody against human pedf  (XpressBio)

Journal: Neoplasia (New York, N.Y.)

Article Title: Pigment Epithelium-Derived Factor Stimulates Tumor Macrophage Recruitment and Is Downregulated by the Prostate Tumor Microenvironment 1

doi:

Figure Lengend Snippet: (A) Western blot analysis of PEDF in PEDF-transfected (MatLyLu-PEDF) and vector-transfected MatLyLu (MatLyLu-CON) cells in vitro. (B) PEDF purified from MatLyLu-PEDF medium was tested for its ability to inhibit migration of HUVECs toward angiogenic FGF2. Results are presented as percentage maximum migration induced by the positive control (FGF2) after the background migration of endothelial cells was subtracted. (C) Cell viability was measured in MatLyLu-PEDF and control cells after 72 hours using an MTT assay. (D) Sections from orthotopic MatLyLu-PEDF and MatLyLu-CON tumors immunostained with an antibody against human PEDF (Chemicon).

Article Snippet: Immunohistochemical staining was performed using primary antibodies against human PEDF (1:100, cat. MAB1059; Chemicon, Temecula, CA), rat TNFα (1:1000, cat. AAR33; AbD Serotec, Oxford, UK), synaptophysin (1:100, cat. A0010; Dako, Stockholm, Sweden), LYVE-1 (1:100, cat. Ab14917; Abcam, Cambridge, UK), or iNOS (1:500, cat. Ab15323; Abcam) incubated overnight.

Techniques: Western Blot, Transfection, Plasmid Preparation, In Vitro, Purification, Migration, Positive Control, MTT Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: Pigment Epithelium-Derived Factor Stimulates Tumor Macrophage Recruitment and Is Downregulated by the Prostate Tumor Microenvironment 1

doi:

Figure Lengend Snippet: (A) Western blot analysis of PEDF in conditioned medium of AT-1 cells in vitro and in tumor lysates from orthotopic AT-1 tumors at day 10. (B) Relative PEDF mRNA levels in AT-1 cells in vitro (n = 3 different batches) and in orthotopic AT-1 tumors in vivo at day 10 (n = 5). (C) Sections from an AT-1 tumor at day 10 showing consecutive areas stained for macrophages (CD68) and TNFα. (D) Relative PEDF mRNA expression in AT-1 cells stimulated with indicated concentrations of TNFα for 18 hours in vitro. (E) Western blot analysis of PEDF in conditioned medium of AT-1 cells stimulated with indicated concentrations of TNFα for 24 hours. (F) Relative PEDF mRNA levels quantified from ventral prostate of Copenhagen rats injected with control (PBS, n = 5) or recombinant rat TNFα (1 µg, n = 5). Values are means ± SD. *P < .05, **P < .01.

Article Snippet: Immunohistochemical staining was performed using primary antibodies against human PEDF (1:100, cat. MAB1059; Chemicon, Temecula, CA), rat TNFα (1:1000, cat. AAR33; AbD Serotec, Oxford, UK), synaptophysin (1:100, cat. A0010; Dako, Stockholm, Sweden), LYVE-1 (1:100, cat. Ab14917; Abcam, Cambridge, UK), or iNOS (1:500, cat. Ab15323; Abcam) incubated overnight.

Techniques: Western Blot, In Vitro, In Vivo, Staining, Expressing, Injection, Recombinant

Journal:

Article Title: Expression of pigment epithelium derived factor and vascular endothelial growth factor in choroidal neovascular membranes and polypoidal choroidal vasculopathy

doi: 10.1136/bjo.2003.032466

Figure Lengend Snippet: An active CNVM (case 1). (A) Preoperative fundus photograph. Subretinal haemorrhage and a serous retinal detachment can be seen. The yellow arrow points to the direction of the sectional OCT scan (C). (B) Preoperative fluorescein angiogram. Hyperfluorescein leakages from the CNVM can be seen. OCT of (A). (C) OCT shows an elevation of the sensory retina by a highly reflective mass above the original RPE layer. (D) Haematoxylin and eosin section showing prominent neovascular vessels. RPE cells (arrows) are observed on the choroidal side. (E) A high power photograph of the region outlined by the black box in (D). Numerous neovascular vessels (arrowheads) are present. (F) Immunohistochemistry for PEDF expression. Strong immunoreactivity for PEDF is observed in endothelial cells (arrowheads) within the neovascularisation and the RPE cells (arrows) located in the perivascular areas. (G) A high power photograph of the region outlined by the black box in (F). (H) Immunohistochemistry for VEGF expression. Strong immunoreactivity for VEGF is observed in endothelial cells (arrowheads) within the neovascularisation and the RPE cells (arrows). (I) A high power photograph of the region outlined by the black box in (H). Scale bars, 50 µm.

Article Snippet: Then, affinity purified rabbit polyclonal primary antibody against human PEDF (BioProducts, Maryland, Inc, MD, USA, 1:300), or affinity purified rabbit polyclonal antibody against human VEGF (Cat No VEGF (A-20):sc-152, Santa Cruz Biotechnology, Inc, CA, USA, 1:200), was applied to sections for 60 minutes followed by incubation with biotinylated goat anti-rabbit IgG.

Techniques: Immunohistochemistry, Expressing

Journal:

Article Title: Expression of pigment epithelium derived factor and vascular endothelial growth factor in choroidal neovascular membranes and polypoidal choroidal vasculopathy

doi: 10.1136/bjo.2003.032466

Figure Lengend Snippet: A quiescent CNVM (case 5). (A) Preoperative fundus photograph and OCT. Subretinal haemorrhage and fibrosis can be seen. The yellow arrow points to the direction of the sectional scan of OCT. OCT shows a thickening of a highly reflective layer corresponding to the RPE and choriocapillaris. High reflective layer consisting of red elements suggest fibrosis. (B) Preoperative fluorescein angiogram. No fluorescein leakage from the CNVM is observed. (C) Preoperative ICG angiogram. (D) ICG angiogram shows tissue staining in the CNVM with a dark rim. Haematoxylin and eosin section. (E) High power photograph of the region outlined by the black box in (D). Fibrosis and proliferation of RPE cells (arrows) are predominant over neovascular vessels (arrowheads). (F) Immunohistochemistry for PEDF expression. Strong immunoreactivity for PEDF is observed in the RPE cells (arrows) located in the perivascular area and the RPE cell layers covering the CNVMs. (G) A high power photograph of the region outlined by the black box in (F). Strong immunoreactivity for PEDF is observed in the RPE cells (arrows) but it is barely detectable in new vessels (arrowheads). (H) Immunohistochemistry for VEGF expression. Immunoreactivity for VEGF is observed in the RPE cells (arrows) located in the perivascular area and RPE cells covering the CNVM, whereas immunoreactivity for VEGF is weak or not detectable in the new vessels (arrowheads). (I) A high power photograph of the region outlined by the black box in (H). Scale bars, 50 µm.

Article Snippet: Then, affinity purified rabbit polyclonal primary antibody against human PEDF (BioProducts, Maryland, Inc, MD, USA, 1:300), or affinity purified rabbit polyclonal antibody against human VEGF (Cat No VEGF (A-20):sc-152, Santa Cruz Biotechnology, Inc, CA, USA, 1:200), was applied to sections for 60 minutes followed by incubation with biotinylated goat anti-rabbit IgG.

Techniques: Staining, Immunohistochemistry, Expressing

Journal:

Article Title: Expression of pigment epithelium derived factor and vascular endothelial growth factor in choroidal neovascular membranes and polypoidal choroidal vasculopathy

doi: 10.1136/bjo.2003.032466

Figure Lengend Snippet: A case of PCV. (A) Preoperative fundus photograph and OCT. Subretinal haemorrhage and retinal pigment epithelium detachment (PED) are observed. The yellow arrow points to the direction of the sectional scan of OCT. OCT shows an elevation of the sensory retina by a highly reflective, dome-like layer. (B) Preoperative fluorescein angiogram displaying fluorescein leakage from the lesion and fluorescein pooling corresponding to the PED. (C) Preoperative ICG angiogram. (D) ICG angiogram displays polyp-like lesions with an associated abnormal vascular network. Haematoxylin and eosin section. The fibrovascular tissue that appears to correspond to the polypoidal lesions. The fibrovascular tissue is observed beneath the dome-like elevation of the RPE layer (arrows). (E) A high power photograph of the region outlined by the black box 1 in (D). The fibrovascular tissue contains numerous dilated thin wall vessels and massive fibrin-like materials. (F) A high power photograph of the region outlined by the black box 2 in (D). The abnormal vessels are surrounded by fibrin-like materials. (G) Immunohistochemistry for PEDF expression. (H) A high power photograph of (G). Strong immunoreactivity for PEDF is observed in the RPE cells (arrows) located above the fibrovascular membrane and also detected in the endothelial cells of the numerous abnormal vessels (arrowheads). (I) Immunohistochemistry for VEGF expression. (J) A high power photograph of (I). Strong immunoreactivity for VEGF is observed in the RPE cells located above the fibrovascular membrane and also in the endothelial cells of the numerous abnormal vessels. Scale bars, 100 µm.

Article Snippet: Then, affinity purified rabbit polyclonal primary antibody against human PEDF (BioProducts, Maryland, Inc, MD, USA, 1:300), or affinity purified rabbit polyclonal antibody against human VEGF (Cat No VEGF (A-20):sc-152, Santa Cruz Biotechnology, Inc, CA, USA, 1:200), was applied to sections for 60 minutes followed by incubation with biotinylated goat anti-rabbit IgG.

Techniques: Immunohistochemistry, Expressing, Membrane