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Ve cadherin

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VE-cadherin is a cell adhesion protein that is primarily expressed in vascular endothelial cells. It plays a key role in the formation and maintenance of cell-cell junctions, which are essential for the integrity and barrier function of the endothelium.

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Lab products found in correlation

Gapdh, by Cell Signaling Technology (9 mentions) β catenin, by Cell Signaling Technology (7 mentions) Smad2 3, by Cell Signaling Technology (5 mentions) β actin, by Cell Signaling Technology (5 mentions) Vegfr2, by Cell Signaling Technology (5 mentions) Sm22α, by Abcam (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Vimentin, by Cell Signaling Technology (4 mentions) Occludin, by Abcam (4 mentions) α sma, by Abcam (4 mentions) Collagen 4, by Abcam (4 mentions) Ve cadherin, by Santa Cruz Biotechnology (3 mentions) N cadherin, by Cell Signaling Technology (3 mentions) Odyssey infrared imaging system, by LI COR (3 mentions) Ab5694, by Abcam (3 mentions) Lsm 780, by Zeiss (3 mentions) Dc protein assay, by Bio-Rad (3 mentions) Claudin 5, by Thermo Fisher Scientific (3 mentions) Phospho smad2, by Cell Signaling Technology (3 mentions) Gapdh, by Merck Group (3 mentions)

Spelling variants of this product

Anti-VE-cadherin (35 citations) Rabbit anti-VE-cadherin (14 citations) VE-cadherin antibody (14 citations) Anti-VE-cadherin antibody (5 citations) Rabbit anti-human VE-cadherin (5 citations) Anti–human VE-cadherin antibody (2 citations) VE-Cadherin (D87F2, (2 citations) Rabbit anti-VE cadherin antibody (2 citations) VE-cadherin XPTM Rabbit antibody (2 citations) VE-cadherin (D87F2) XP (2 citations)

98 protocols using ve cadherin

1

Immunocytochemistry and Immunohistochemistry of Cadherins

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Rabbit monoclonal E-cadherin antibody serum (Cell Signaling Technology, Inc, Danvers, MA, USA), which recognizes the human E-cadherin epitope, was used for immunocytochemistry at a 1:200 dilution and for immunohistochemistry using a 1:400 dilution according to the manufacturer’s instructions. Purified rabbit monoclonal N-cadherin (Cell Signaling Technology, Inc) and VE-cadherin (Cell Signaling Technology, Inc) antibody serum, which recognizes the human N-cadherin and VE-cadherin epitopes, respectively, were used in immunocytochemistry at a 1:200 dilution according to the manufacturer’s instructions.
DyLight™ 488 donkey anti-rabbit IgG secondary antibody (Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) was used for immunocytochemistry to detect E-cadherin and VE-cadherin primary antibodies. A final concentration of 40 μg/mL was used. Alexa Fluor® 594 goat anti-rabbit secondary antibody (Santa Cruz Biotechnology, Inc) was used for immunocytochemistry to detect N-cadherin primary antibodies at a final concentration of 1 μg/mL. For immunohistochemistry, polyclonal goat anti-rabbit biotinylated immunoglobulin (Dako Canada, Inc, Burlington, ON, Canada) was used as a secondary antibody at a final concentration of 1.9 μg/mL.
O’Shea L.K., Abdulkhalek S., Allison S., Neufeld R.J, & Szewczuk M.R. (2014). Therapeutic targeting of Neu1 sialidase with oseltamivir phosphate (Tamiflu®) disables cancer cell survival in human pancreatic cancer with acquired chemoresistance. OncoTargets and therapy, 7, 117-134.
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2

Protein Expression Analysis in HACECs

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HACECs were re-suspended in lysis buffer and 2 mg/ml protease inhibitor cocktail (Roche Diagnostics Corp). Protein concentrations were determined using the Bradford Protein assay kit (Beyotime Biotechnology, China). An equal amount of protein was analyzed by SDS-PAGE and immunoblotting. Primary antibodies used included the following: α-SMA (ab5694, abcam, USA), calponin (ab46794, abcam, USA), VE-cadherin (Cat. 2500, Cell Signaling Technology, USA), PECAM1 (sc-71872, Santa Cruz, USA), SMURF2 (GTX31571, GeneTex, USA) and GAPDH (ab8245, abcam, USA). Secondary antibodies were fluorescence-labeled antibodies. Bands were visualized using the Odyssey Infrared Imaging System (LI-COR Biotechnology).
Lou C, & Li T. (2020). Long non-coding RNA SENCR alleviates endothelial-to-mesenchymal transition via targeting miR-126a. Archives of Medical Science : AMS, 19(1), 180-188.
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3

Immunofluorescence Analysis of Endothelial Junctions

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For immunofluorescence studies, HMVEC-L and HPAEC cells were cultured on collagen/fibronectin-coated filter inserts. Endothelial monolayers were fixed using a mixture of ice-cold ethanol:acetic acid (95:5 v/v) for 10 min and then washed three times for 5 min in PBS. After blocking with 3% bovine serum albumin (BSA, Sigma) for 30 min, inserts were incubated with primary antibodies against VE-cadherin (Cell Signaling Technology) or ZO-1 (Invitrogen), nuclei were counterstained with Hoechst33342 (Sigma). The staining was visualized using Cy3- or Cy2-conjugated secondary antibodies (Jackson Immuno Research) diluted in 1% BSA-containing PBS, and washed three times for 5 min in PBS. Filter inserts were mounted in anti-fading embedding medium (Fluorogel, Electron Microscopy Sciences, Hatfield) and studied using a Nikon Eclipse TE2000U photomicroscope (Tokyo, Japan) connected to a digital camera (Spot RT KE, Diagnostic Instruments, Sterling Heights, MI, United States).
Immunofluorescence images were quantified using the ImageJ software (version 1.51n, NIH). We measured the mean intensity of VE-cadherin immunofluorescence staining using the polygon selection tool to define the cell junctions. In the case of ZO-1 tight junctional protein, we selected the cells by freehand selection tools and then measured the continuity of the immunofluorescence staining.
Fazakas C., Nagaraj C., Zabini D., Végh A.G., Marsh L.M., Wilhelm I., Krizbai I.A., Olschewski H., Olschewski A, & Bálint Z. (2018). Rho-Kinase Inhibition Ameliorates Dasatinib-Induced Endothelial Dysfunction and Pulmonary Hypertension. Frontiers in Physiology, 9, 537.
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4

Immunofluorescence Staining of Cardiac Cells

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Cells were fixed with 4% PFA and stained with monoclonal anti-α-Actinin (Sarcomeric) (A7811, Sigma), VE-cadherin (2500, Cell Signaling Technology), PECAM-1 (M0823, Dako), MYL7 (17283-1-AP, Proteintech), MYL4 (67533-1-Ig, Proteintech) and cTnT (MAB1874, R&D Systems) antibodies. Nuclei were visualized with DAPI. Stained cells were imaged with fluorescent or confocal microscopes (Zeiss AxioImager and Zeiss LSM 780).
Helle E., Ampuja M., Dainis A., Antola L., Temmes E., Tolvanen E., Mervaala E, & Kivelä R. (2021). HiPS-Endothelial Cells Acquire Cardiac Endothelial Phenotype in Co-culture With hiPS-Cardiomyocytes. Frontiers in Cell and Developmental Biology, 9, 715093.
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5

Quantifying Wnt Signaling Pathway

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Primary antibodies used were total β-catenin (Cat No. 2677; 1:100), active (non-phosphorylated) β-catenin (Cat No. 8814; 1:100), VE-Cadherin (Cat No.2500; 1:100) (Cell Signaling, Danvers, MA). For full details see online methods.
Hox V., O’Connell M.P., Lyons J.J., Sackstein P., Dimaggio T., Jones N., Nelson C., Boehm M., Holland S.M., Freeman A.F., Tweardy D.J., Olivera A., Metcalfe D.D, & Milner J.D. (2016). Diminution of STAT3 signaling inhibits vascular permeability and anaphylaxis. The Journal of allergy and clinical immunology, 138(1), 187-199.
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6

Immunofluorescence Imaging of Cell Markers

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For immunofluorescence staining, the cells were incubated with primary antibodies against VE-cadherin (Cell Signaling Technology [CST], USA), vimentin (CST) or fibronectin (Abcam), followed by incubation with Alexa Fluor 488- or 555-conjugated secondary antibodies (Invitrogen, USA). For confocal microscopy, the cells grown on coverslips were counterstained with DAPI and were imaged using a confocal laser-scanning microscope (Carl Zeiss, Germany) with a core data acquisition system (Applied Precision).
Lin L., Chen Y.S., Yao Y.D., Chen J.Q., Chen J.N., Huang S.Y., Zeng Y.J., Yao H.R., Zeng S.H., Fu Y.S, & Song E.W. (2015). CCL18 from tumor-associated macrophages promotes angiogenesis in breast cancer. Oncotarget, 6(33), 34758-34773.
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7

Western Blot Analysis of Cell Signaling Proteins

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After adjusting the total protein concentration of the cell samples, 50 μg of protein sample was uploaded per well and separated by polyacrylamide gel electrophoresis. The proteins were transferred to PVDF membrane (cat. no. FFP24; Beyotime) and then washed with PBS (cat. no. C0221A; Beyotime) for three times. After 10 min each, the membrane was blocked with 5% skim milk (cat. no. P0216; Beyotime) at room temperature for 2 h. Next, the membrane was probed with the corresponding primary antibodies at 4°C for incubation overnight and then incubated with secondary antibody Goat Anti‐Rabbit IgG H&L (HRP; ab6721; dilution, 1:2000; Abcam) at 37°C for 2 h. After washing with TBST again, it was placed into Odyssey (LI‐COR) for scanning and analysis. The primary antibodies included P2Y12R (ab233760; dilution, 1:1000; Abcam), NF‐κB p65 (ab32536; dilution, 1:1000; Abcam), P‐IKB‐α (ab133462; dilution, 1:10,000; Abcam), IKB‐α (ab32518; dilution, 1:1000; Abcam), p‐AKT (ab38449; dilution, 1:1000; Abcam), eNOS (ab300071; dilution, 1:1000; Abcam), VE‐cadherin (#2500; dilution, 1:1000; Cell Signaling Technology), ZO‐1 (ab216880; dilution, 1:1000; Abcam), Claudin 5 (ab131259; dilution, 1:1000; Abcam), Occludin (ab216327; dilution, 1:1000; Abcam), H3 (ab1791; dilution, 1:1000; Abcam), and GAPDH (ab8245; dilution, 1:1000; Abcam).
, & Han X. (2022). Inhibiting P2Y12 receptor relieves LPS‐induced inflammation and endothelial dysfunction. Immunity, Inflammation and Disease, 10(10), e697.
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8

Immunoblotting Assay for Protein Expression

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Cells were lysed in SDS buffer (60mM TrisHCl pH 6.8, 5% glycerol, 2% SDS). Twenty-five to 50 μg of cell lysates were run on SDS-polyacrylamide gels, transferred to PVDF membranes (Millipore, Billerica, MA), and incubated 1 hour in a blocking buffer (5% Bovine Serum Albumin (BSA), 10 mM Tris–HCl, pH 7.6, 150 mM NaCl, and 0.1%Tween 20). Each membrane was incubated overnight against a specific antibody: SMAD2 (Cell Signaling), pSMAD2 (Cell Signaling), SMAD1 (Cell Signaling), SMAD5 (Cell Signaling), pSMAD1/5 (Cell Signaling), PAI-1(Santa Cruz Inc., Dallas, TX), VE-cadherin (Cell Signaling), Calponin (Millipore), PECAM1 (Abcam), SM22α (Abcam). After incubation with peroxidase conjugated secondary antibodies, detection was performed with enhanced chemiluminescence reagents (GE Healthcare, Sweden). Protein levels of GAPDH (R&D Systems, Minneapolis, MN) were used to normalize the results.
Miyakawa A.A., Girao-Silva T., Krieger J.E, & Edelman E.R. (2018). RAPAMYCIN ACTIVATES TGF RECEPTOR INDEPENDENTLY OF ITS LIGAND: IMPLICATIONS FOR ENDOTHELIAL DYSFUNCTION.. Clinical science (London, England : 1979), 132(4), 437-447.
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9

Immunofluorescence Analysis of HUVEC Junctions

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HUVECs were seeded on coverslips and incubated in the presence of MAPF for 8 h. Cells were then rinsed with PBS and fixed with 10% v/v formalin in PBS (pH 7.4). A blocking solution (5% milk in 0.1% v/v Triton X-100) was applied to prevent nonspecific binding. Primary antibody against VE-cadherin (Cell Signaling Technology, USA) and β-catenin (BD Bioscience, USA) in blocking buffer was incubated with the HUVECs at 4 °C overnight. After the antibody was washed, the slides were incubated with fluorescein isothiocyanate-conjugated goat anti-mouse and anti-rabbit IgG (Sigma-Aldrich) for 1 h. Finally, coverslips were mounted with mounting medium (Gel Mount Aqueous, Sigma) and photographed with a Nikon D1X digital camera (Carl Zeiss, Oberkochen, Germany).
Chen W.T., Lin Y.H., Changchien C.Y., Chen Y., Chang H.H., Tsai W.C., Tsai H.C., Wang C.Y., Shen M.S., Cheng L.T, & Tsai C.L. (2021). Concurrent Blockade of Endothelial EGFR and VEGF Signaling on Malignant Associated Pleural Fluid Induced Angiogenesis: From Clinic to Bench. Biomedicines, 9(10), 1327.
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10

Immunoblotting of Signaling Pathways in Cell Lysates

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The cell lysates were prepared using immunoprecipitation assay lysis buffer (Millipore, Burlington, MA, USA) supplemented with protease and phosphatase inhibitors (Roche, Basel, Switzerland). Protein concentration was measured by the DC protein assay (Bio-Rad, Hercules, CA, USA), and approximately 30–40 μg of proteins in Laemmli buffer were used. Western blotting was performed as described previously [29 (link)]. Densitometry was performed using NIH ImageJ software. Antibodies used include pSer473-Akt (Cat No. 9271), pThr308-Akt (Cat No. 9275), Pan Akt (Cat No. 4685), pP38 MAP Kinase (Cat No. 9215), total P38 MAP Kinase (Cat No. 9212), VE-cadherin (Cat No. 2500), and GAPDH (Cat No. 5174), were purchased from Cell Signaling, Danvers, MA, USA. Anti-Claudin-5 (CLDN5) (Cat No. 352500) antibody was purchased from Thermo Scientific, Waltham, MA, USA. Anti-mouse (Cat No. 170-6516) and anti-rabbit (Cat No. 170-6515) HRP-conjugated secondary antibodies were obtained from Bio-Rad (Hercules, CA, USA). Alexa 485-conjugated secondary antibodies were purchased from Thermo Scientific (Frederick, MD, USA). Antibody dilutions are provided in Appendix A.
Rudraraju M., Narayanan S.P, & Somanath P.R. (2021). Distinct Mechanisms of Human Retinal Endothelial Barrier Modulation In Vitro by Mediators of Diabetes and Uveitis. Life, 12(1), 33.
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