Rat anti ve cadherin

Manufactured by BD

Rat anti-VE-cadherin is a laboratory reagent used for the detection and analysis of VE-cadherin, a cell adhesion protein found in endothelial cells. It can be used in various applications such as flow cytometry, immunohistochemistry, and Western blotting to study the expression and localization of VE-cadherin.

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Close spelling variants of this product

Rat anti–mouse VE-Cadherin Anti-VE-cadherin VE-cadherin

4 protocols using rat anti ve cadherin

Retinal Vessel Imaging in Tecr Mice

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Retinas were harvested from WT and Tecr^iECKO mice as described previously [57 (link)]. In brief, mice were perfused with phosphate-buffered saline (PBS) and 4% paraformaldehyde (PFA). Tissue was fixed for 15 min at room temperature and washed with PBS. Whole-mount retinas were dissected to isolate the retinal cup, and relieving cuts were made prior to staining. Brains were fixed by immersion in 4% PFA/PBS overnight at 4°C, dehydrated in 30% sucrose, and then frozen in TissueTek OCT (Sakura). After fixation, retinas or brain sections were blocked for 2 h in 10% goat serum/5% BSA with 0.5% Triton X-100 and incubated overnight with the following antibodies: isolectin B4, Alexa Fluor 647-conjugate (Thermo Fisher Scientific, 1 : 100), rat anti-VE-cadherin (BD Biosciences, 1 : 250), rat anti-CD31 (Abcam, 1 : 200), rat anti-Tecr (Thermo Fisher Scientific, 1 : 100), rat anti-Plvap (BD Biosciences, 1 : 200), mouse anti-Claudin-5, and Alexa Fluor 488-conjugated (Thermo Fisher Scientific, 1 : 200). For detection, suitable species-specific Alexa Fluor-coupled secondary antibodies (1 : 500) were used. Tissues were imaged using confocal microscopy (Leica).

Wang J., Xu J., Zang G., Zhang T., Wu Q., Zhang H., Chen Y., Wang Y., Qin W., Zhao S., Qin E., Qiu J., Zhang X., Wen L., Wang Y, & Wang G. (2022). trans-2-Enoyl-CoA Reductase Tecr-Driven Lipid Metabolism in Endothelial Cells Protects against Transcytosis to Maintain Blood-Brain Barrier Homeostasis. Research, 2022, 9839368.

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Recombinant Protein and Antibody Analysis

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Recombinant human VEGF₁₆₅ (Ala27-Arg191, no. 293-VE), human Sema3C (Gly21-Ser738, no. 5570-S3), and mouse Sema3C (Gln24-Ser741, no. 1728-S3) were from R&D Systems. Recombinant human IgG-Fc (009-000-008) was purchased from Jackson ImmunoResearch. Antibodies: rat anti-Sema3C (clone 238835) and goat anti-Nrp-1 (AF566) are from R&D Systems; rabbit anti-pY397FAK, rabbit anti-pT180/pY182-p38MAPK (Invitrogen); rabbit anti-FAK (C-20), and goat anti-Nrp-1 (C-19) are from Santa Cruz; mouse anti-p38MAPK (L53F8), rabbit anti-AKT, rabbit anti-pS473AKT, and rabbit anti-cleaved Caspase-3 (Asp175) are from Cell Signaling; pTyr118 paxillin (NEB), rat anti-CD34 (NCL-END; Novocastra); rat anti-CD31 (clone Mec13.3) and rat anti-VE-cadherin are from BD Biosciences; mouse anti-vinculin (V9131) and anti-actin smooth muscle-Cy3 are from Sigma; rabbit anti-GFAP (DAKO), rabbit anti-NG2 (Millipore), rabbit anti-collagen IV (Cosmo Bio).

Yang W.J., Hu J., Uemura A., Tetzlaff F., Augustin H.G, & Fischer A. (2015). Semaphorin-3C signals through Neuropilin-1 and PlexinD1 receptors to inhibit pathological angiogenesis. EMBO Molecular Medicine, 7(10), 1267-1284.

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Immunostaining of Extracellular Matrix Proteins

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En face mounted tissue samples were rehydrated with water for 10 minutes. The tissue was then permeabilized using Tris-Triton buffer (50 mM Tris, 500 mM NaCl, 0.3% Triton X-100) for 1 hour, followed by 3 washes with water. Permeabilized tissue was then blocked using 0.5% non-fat dry milk and 1% BSA in TBST for 60 minutes at room temperature. Primary antibodies (Rabbit anti-FN, 1:100, Sigma F3648, St. Louis, MO; rat anti-VE-Cadherin, 1:50, BD Pharmingen 550548, Franklin Lakes, NJ,; or rabbit anti-Collagen α1, 1:100, Abcam Ab292, Cambridge, UK) were then diluted in blocking buffer and incubated with the tissue overnight at 4 °C. After primary antibody incubation, slides were washed 3 times with TBST. Secondary antibodies (Goat anti-rat Alexafluor 488, 1:100, A11006 Life Technologies; Goat anti-rabbit Alexafluor 568, 1:100, A11011 Life Technologies) were diluted in blocking buffer and incubated with the tissue for 60 minutes at room temperature. The tissue was washed 3 times in TBST, and then mounted with VectaShield plus DAPI (Vector Labs, Burlingame, CA) and sealed with clear nail polish. Slides were imaged on a Zeiss 700 or Zeiss 710 confocal microscope.

Chen Z., Givens C., Reader J.S, & Tzima E. (2017). Haemodynamics Regulate Fibronectin Assembly via PECAM. Scientific Reports, 7, 41223.

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En Face Staining of Mouse Aorta

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En face staining was performed as previously described in detail 43 (link), with minor modifications 42 (link), 44 (link). Mouse aortas were collected after perfusion with saline and neutrally buffered 4% PFA. Aortic segments were then permeabilized with 0.1% Triton X-100 in PBS for 10 min and blocked with 10% normal goat serum (Invitrogen) in Tris-buffered saline (TBS) containing 2.5% Tween-20 for 30 min at room temperature. Next, aorta segments were incubated with rat anti-VE-Cadherin (1:100; #555289, BD Bioscicence), rabbit anti-EZH2 (#6263, ProSci) or rabbit anti-H3K27me3 (#39155, Active Motif) antibody overnight at 4 °C. After rinsing with washing buffer 3 times, aortic segments were incubated with Alexa Fluor 488-conjugated goat anti-rat and Alexa Fluor 546-conjugated goat anti-rabbit secondary antibodies (1:1,000; Thermo Fisher Scientific) for 1 h in darkness at room temperature. Finally, aortic segments were carefully whole mounted (with lumen side up) in the ProLong Gold-antifade Mounting Media (Invitrogen) for confocal microscopy (IX81, Olympus) with 60x or 40x oil lens. All animal procedures conformed to the Guideline for the Care and Use of Laboratory Animals published by the U.S. National Institutes of Health and were approved by the Institutional Animal Care and Use Committee of the University of Rochester Medical Center.

Xu S., Xu Y., Yin M., Zhang S., Liu P., Koroleva M., Si S., Little P.J., Pelisek J, & Jin Z.G. (2018). Flow-dependent epigenetic regulation of IGFBP5 expression by H3K27me3 contributes to endothelial anti-inflammatory effects. Theranostics, 8(11), 3007-3021.

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