Goat anti human vegf r3

Manufactured by R&D Systems

Goat-anti-human VEGF-R3 is an affinity-purified antibody that recognizes the human vascular endothelial growth factor receptor 3 (VEGF-R3) protein. VEGF-R3 is a receptor tyrosine kinase that plays a role in lymphangiogenesis and angiogenesis.

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Lab products found in correlation

Goat anti human lyve 1, by R&D Systems (1 mentions) Goat anti human vimentin, by R&D Systems (1 mentions) Goat anti mouse vegfr3, by R&D Systems (1 mentions) Af488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Duolink in situ reagents, by Olink (1 mentions) Horseradish peroxidase labeled secondary antibody, by Agilent Technologies (1 mentions) Immobilon p pvdf, by Merck Group (1 mentions) Mouse anti hsc70 sc 7298, by Santa Cruz Biotechnology (1 mentions) Af349, by R&D Systems (1 mentions) Mini protean tgx precast gel, by Bio-Rad (1 mentions)

3 protocols using goat anti human vegf r3

TGF-β Modulation of Lymphatic Markers

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LECs were treated with human TGF-β1, -β2 and β3 for the indicated time points. Lysates were prepared and subjected to Western blot analysis using 5 μg/ml polyclonal rabbit-anti-human Prox-1 (Reliatech, Cat. No. 102-PA32AG), 1 μg/ml polyclonal goat-anti-human Lyve-1 (R&D Systems, Cat. No. AF2089), 0.2 μg/ml polyclonal goat-anti-human VEGF-R3 (R&D Systems, Cat. No. AF349) and 0.1 μg/ml polyclonal goat-anti-human vimentin (R&D Systems, Cat. No. AF2105) antibodies. For evaluation of protein loading, the blot was probed with 0.01 μg/ml monoclonal mouse-anti-human vinculin-specific antibodies (Sigma Aldrich, Taufkirchen, Germany, Cat. No. V9264). For densitometric evaluation, bands for the Prox-1, Lyve-1, vimentin and VEGFR-3 proteins were normalized to the corresponding loading control using the software ImageJ. Values are presented relative to untreated control samples.

Scherer S.D., Bauer J., Schmaus A., Neumaier C., Herskind C., Veldwijk M.R., Wenz F, & Sleeman J.P. (2016). TGF-β1 Is Present at High Levels in Wound Fluid from Breast Cancer Patients Immediately Post-Surgery, and Is Not Increased by Intraoperative Radiation Therapy (IORT). PLoS ONE, 11(9), e0162221.

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Proximity Ligation Assay for VEGFR Phosphorylation

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Proximity ligation assays were performed to determine tyrosine phosphorylation of VEGFRs or protein interactions (Soderberg et al, 2006) according to the manufacturer's protocol using the Duolink in situ reagents (Olink Bioscience). The following antibodies were used on embryonic sections: goat anti‐mouse VEGFR3 (R&D Systems, AF743), rabbit anti‐mouse VEGFR2 clone 55B11 (Cell Signaling, 2479), mouse anti‐phospho‐tyrosine 4G10 Platinum (Millipore, 05‐1050) and rat anti‐mouse β1 integrin clone MB1.2 (Millipore, MAB1997). The following antibodies were used for in vitro PLA studies: goat anti‐human VEGFR3 (R&D Systems, AF349) and mouse anti‐human β1 integrin (Millipore, MAB1987). Co‐stainings were performed using rabbit anti‐mouse Lyve‐1 (Abcam, ab14917) or Alexa Fluor^®488 phalloidin (Invitrogen by Thermo Fisher, A12379). Secondary antibodies conjugated with AF488 (Molecular Probes) were used, as well as DAPI (Sigma) to counterstain cell nuclei. All images were acquired using Laser Scan Microscopy (LSM 710, Zeiss) and analysed using the ImageJ software. PLA dots were normalised to the number of LECs or to the Lyve1‐positive area.

Urner S., Planas‐Paz L., Hilger L.S., Henning C., Branopolski A., Kelly‐Goss M., Stanczuk L., Pitter B., Montanez E., Peirce S.M., Mäkinen T, & Lammert E. (2018). Identification of ILK as a critical regulator of VEGFR3 signalling and lymphatic vascular growth. The EMBO Journal, 38(2), e99322.

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Quantification of VEGFR2 and VEGFR3 Expressions

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Lysates containing equal amounts of protein from lungs were separated in 7.5% Mini-PROTEAN TGX Precast gels (BIORAD). After blotting to polyvinylidene fluoride membranes (Immobilon-P PVDF; Millipore), the proteins were detected using goat anti-mouse VEGFR2 (AF644, 1:1000; R&D Systems), goat anti-mouse VEGFR3 (AF743, 1:1000; R&D Systems), mouse anti-HSC-70 (SC-7298, 1:5000; Santa Cruz Biotechnology), rabbit anti-human pVEGFR2 Tyr1175 (19A10, 1:1000; Cell Signaling), goat anti-human VEGFR2 (AF357, 1:500; R&D Systems), goat anti-human VEGFR3 (AF349, 1:1000; R&D Systems), or mouse anti-human VEGFR3 (9D9,^{25 (link)} 1:1000)–specific primary antibodies. The blots were then probed with horseradish peroxidase–labeled secondary antibodies (Dako, Glostrup, Denmark), and the signal was visualized with the SuperSignal West Pico Chemiluminescent Substrate or the SuperSignal West Femto Maximum Sensitivity Substrate (Pierce, Rockford, IL).

Heinolainen K., Karaman S., D’Amico G., Tammela T., Sormunen R., Eklund L., Alitalo K, & Zarkada G. (2017). VEGFR3 Modulates Vascular Permeability by Controlling VEGF/VEGFR2 Signaling. Circulation research, 120(9), 1414-1425.

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