Rat VEGF Antibody

VEGF₁₆₄in Rat Kidney. VEGF₁₆₄was detected in perfusion fixed frozen sections of rat kidney using 15 µg/mL Goat Anti-Rat VEGF₁₆₄Antigen Affinity-purified Polyclonal Antibody (Catalog # AF564) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Cell Proliferation Induced by VEGF₁₆₄and Neutralization by Rat VEGF Antibody. Recombinant Rat VEGF₁₆₄(Catalog # 564-RV) stimulates proliferation in HUVEC human umbilical vein endothelial cells in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat VEGF₁₆₄(20 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat VEGF 164 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF564). The ND₅₀ is typically 0.2-0.6 µg/mL.

Detection of Recombinant Rat and Mouse VEGF by Western Blot. Western blot shows 25 ng of Recombinant Rat VEGF 164 (Catalog # 564-RV), Recombinant Human VEGF 165 (Catalog # 293-VE) and Recombinant Mouse VEGF 164 (Catalog # 493-MV). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Rat VEGF 164 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF564) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for VEGF at approximately 20-25 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.This antibody does not detect natural VEGF in lysates from cell lines or tissues.

Detection of Porcine VEGF by Immunohistochemistry Expression of proteins relevant to kidney regeneration in female MWF rats.Immunohistochemical staining of VEGF in female animals receiving saline or MET transplant, after 6 weeks. Scale bars = 50 μm. Representative immunofluorescence staining (red) of FGF2, HGF, IGF-1 and Pax-2 in female rats receiving saline or MET, on renal tissues labeled with WGA-lectin (green) and DAPI (blue). Scale bars = 50 μm, = 25 μm (for FGF2 and IGF-1). Semiquantitative analysis of immunohistochemical stainings of VEGF, FGF2, HGF, IGF-1 and Pax-2 in female MWF rats receiving saline or MET. Data are expressed as mean ± SE (n = 3/group). Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0120235), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Porcine VEGF by Western Blot Depletion of BAMs attenuates ischemia-induced pial and cortical vascular permeability. BAMs were depleted by i.c.v. administration of clodronate liposomes (CL) 4 days prior to ischemia. For treatment control, rats received vehicle liposomes (V). a) Vegfa mRNA was studied in cortical and subcortical regions of the ipsilateral (ipsi, ischemic) and the contralateral (contra) brain hemispheres. Ischemia-induced expression of Vegfa mRNA at 24 h is attenuated in the cortex of the CL rats versus the V rats (n = 6 per group) (Mann Whitney test, p = 0.041). b) Western blotting of cortical and subcortical brain tissue (ipsilateral) 24 h post-ischemia shows the VEGF164 isoform of VEGF-A detected as two bands corresponding to the homodimeric and monomeric forms at approximately 54 kDa and 24 kDa. Quantification of signal intensity of the 54-kDa band shows reduced expression in the cortex of rats receiving CL vs. rats receiving V (n = 6 per group) (Mann-Whitney test, **p = 0.002). c) Ischemia increases the permeability of pial vessels (arrows), as assessed by Evans blue extravasation 24 h post-ischemia, and BAM depletion attenuates the effect. Images of one representative coronal brain section per rat of each treatment group show Evans blue extravasation in the ipsilateral hemisphere. Rats receiving clodronate show negligible Evans blue in the cortex (arrows in the schematic representation at the right hand side). The volume of tissue showing Evans blue extravasation is reduced in the cortex, but not in subcortical zones, of the CL group (n = 8) versus the V group (n = 7) (Mann-Whitney test, *p = 0.014). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30092836), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Porcine VEGF by Immunohistochemistry Expression of mRNAs and proteins relevant to kidney regeneration in male MWF rats.(A) Real Time RT-PCR analysis of VEGF, FGF2, HGF, IGF-1 and Pax-2 in whole host renal tissues of male MWF rats receiving saline or metanephroi (MET) (in adjacent area), 6 weeks after transplantation. Data are expressed as mean ± SE; *P < 0.05, **P < 0.01 vs saline (n = 4 rats/group, VEGF n = 6 rats/group). (B) Immunohistochemical staining of VEGF (arrows) in male animals receiving saline or MET transplant, after 6 weeks. Scale bars = 50 μm. Representative immunofluorescence stainings (red) of FGF2, HGF, IGF-1 and Pax-2 in rats receiving saline or MET, on renal tissues labeled with WGA-lectin (green) and DAPI (blue). Scale bars = 50 μm, = 25 μm (for FGF2 and IGF-1). Semiquantitative analysis of immunohistochemical staining of VEGF, FGF2, HGF, IGF-1 and Pax-2 in male MWF rats receiving saline or MET. Data are expressed as mean ± SE; *P < 0.05, **P < 0.01 vs saline (n = 3/group). Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0120235), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Porcine VEGF by Immunocytochemistry/ Immunofluorescence Effect of fibroblasts transplanted under the kidney capsule of male MWF rats.(A) Histological appearance of renal tissue from male MWF rat transplanted with fibroblasts in adjacent (left, arrow) or distant (right) area. Scale bars = 200 μm. (B) Quantification of endothelium volume density (Vv), peritubular capillary length density (Jv), nitrotyrosine (NT)-positive tubuli, TUNEL-positive cells and Ki-67-positive cells, in renal tissues of MWF rats receiving fibroblasts or saline. Data are expressed as mean ± SE (n = 3 rats/group). (C) Representative immunohistochemical images of SMP30 (green), NCAM (red), VEGF (brown signal), FGF2 (red), HGF (red), IGF-1 (red) and Pax-2 (red) in male rats with fibroblast transplant. Sections are co-stained with rhodamine-LCA (in SMP30 panel, red) or with FITC-WGA lectin (in panels showing NCAM, FGF2, HGF, IGF-1, Pax2, green) and DAPI (blue). Scale bars = 50 μm. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0120235), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rat VEGF by Immunohistochemistry Expression of mRNAs and proteins relevant to kidney regeneration in male MWF rats.(A) Real Time RT-PCR analysis of VEGF, FGF2, HGF, IGF-1 and Pax-2 in whole host renal tissues of male MWF rats receiving saline or metanephroi (MET) (in adjacent area), 6 weeks after transplantation. Data are expressed as mean ± SE; *P < 0.05, **P < 0.01 vs saline (n = 4 rats/group, VEGF n = 6 rats/group). (B) Immunohistochemical staining of VEGF (arrows) in male animals receiving saline or MET transplant, after 6 weeks. Scale bars = 50 μm. Representative immunofluorescence stainings (red) of FGF2, HGF, IGF-1 and Pax-2 in rats receiving saline or MET, on renal tissues labeled with WGA-lectin (green) and DAPI (blue). Scale bars = 50 μm, = 25 μm (for FGF2 and IGF-1). Semiquantitative analysis of immunohistochemical staining of VEGF, FGF2, HGF, IGF-1 and Pax-2 in male MWF rats receiving saline or MET. Data are expressed as mean ± SE; *P < 0.05, **P < 0.01 vs saline (n = 3/group). Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0120235), licensed under a CC-BY license. Not internally tested by R&D Systems.