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Anti ve cadherin ab33168

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Anti-VE-cadherin (ab33168) is a primary antibody that recognizes the VE-cadherin protein. VE-cadherin is a cell-cell adhesion molecule expressed in vascular endothelial cells and is important for the formation and maintenance of the endothelial barrier.

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Lab products found in correlation

Formamide, by Merck Group (2 mentions) Anti ve cadherin f 8, by Santa Cruz Biotechnology (2 mentions) Anti trio h 120, by Santa Cruz Biotechnology (2 mentions) Anti notch1 icd c 20 r, by Santa Cruz Biotechnology (2 mentions) Anti notch1 ecd h 131, by Santa Cruz Biotechnology (2 mentions) Anti ve ptp h 300, by Santa Cruz Biotechnology (2 mentions) Anti rfp ab124754, by Abcam (2 mentions) Af647 conjugate if, by Cell Signaling Technology (2 mentions) Anti rac1 antibody 102, by BD (2 mentions) Rhodamine and alexa fluor 488 labelled phalloidin, by Thermo Fisher Scientific (2 mentions) Anti ptprf antibody sab4200321, by Merck Group (2 mentions) Dynasore hydrate, by Merck Group (2 mentions) Anti β tubulin ab6046, by Abcam (2 mentions) Rac1 inhibitor nsc23766, by Santa Cruz Biotechnology (2 mentions) Evans blue dye, by Merck Group (2 mentions) Anti gapdh ab9485, by Abcam (1 mentions) Facsaria 3 cell sorter, by BD (1 mentions) Duolink in situ pla probe anti mouse minus, by Merck Group (1 mentions) Anti cd31 af3628, by R&D Systems (1 mentions) Phenylmethanesulfonyl fluoride pmsf, by Beyotime (1 mentions)

5 protocols using anti ve cadherin ab33168

1

Co-culture Assay for Brain Metastasis

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Pre-transfected MDA231-BrM2 cells were co-cultured for 24 hours with hCMEC/D3 monolayers, which were stably transduced with a lentivirus expressing a triple-fusion reporter (TGL) encoding firefly luciferase and green fluorescence protein (GFP).29 (link) GFP-expressing cancer cells were separated from unlabeled hCMEC/D3 by fluorescence-activated cell sorting (FACS) using the BD FACSAria™ III cell sorter (BD Biosciences, USA). The collected hCMEC/D3 cells were then lysed for western blot analysis according to standard procedures and as previously described.13 (link),15 (link) The following primary antibodies were used: Anti-Claudin 5 (4C3C2) (35–2500) 1/500 (Thermo Fisher Scientific), Anti-VE-cadherin (ab33168) 1/1000, Anti-MMP-1 (ab38929) 1/5000 (Abcam), and Anti-GAPDH (ab9485) 1/7500 (Abcam) (as loading control).
Hammash D., Mahfood M., Khoder G., Ahmed M., Tlili A., Hamoudi R, & Harati R. (2022). miR-623 Targets Metalloproteinase-1 and Attenuates Extravasation of Brain Metastatic Triple-Negative Breast Cancer Cells. Breast Cancer : Targets and Therapy, 14, 187-198.
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2

Molecular Mechanisms of Endothelial Activation

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Lipopolysaccharide LPS L2880 was obtained from Sigma-Aldrich St. Louis, MO, USA. A myeloperoxidase (MPO) assay kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Anti-CD31 (AF3628) was purchased from R&D Systems (Minneapolis, MN, USA). Anti-MYH9 (11128–1-AP) and anti-IgG (B900610) were purchased from Proteintech Group (Chicago, IL, USA). Anti-Wnt5a (sc-365,370), Anti-β-catenin (sc-7963), Anti-TLR4 (sc293072), and Protein A/G PLUS-Agarose (sc-2003) were obtained from Santa Cruz Biotechnology Inc. (Texas, CA, USA). Anti-VE-cadherin (ab33168) was provided by Abcam (Cambridge, MA, USA). Duolink®In Situ PLA® Probe Anti-Rabbit PLUS (DUO92002), Duolink® In Situ PLA® Probe Anti-Mouse MINUS (DUO92004), and Duolink®In Situ Detection Reagents Red (DUO92008) were obtained from Sigma–Aldrich (St. Louis, MO, USA).
The bicinchoninic acid (BCA) protein assay kit and phenylmethanesulfonyl fluoride (PMSF) were procured from Beyotime Biotechnology (Shanghai, China). Alexa Fluor 488- and 594-conjugated secondary antibodies and Dynabeads™ Sheep Anti-Rat IgG (11035) were from Thermo Fisher Scientific (Waltham, MA, USA).
Zhang J., Pan Z., Zhou J., Zhang L., Tang J., Gong S., Li F., Yu B., Zhang Y, & Kou J. (2022). The myosin II inhibitor, blebbistatin, ameliorates pulmonary endothelial barrier dysfunction in acute lung injury induced by LPS via NMMHC IIA/Wnt5a/β-catenin pathway. Toxicology and Applied Pharmacology, 450, 116132.
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3

Immunofluorescence Staining of VE-Cadherin

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After particle flow (see above), cells were fixed with 4% PFA, washed twice with PBS 1%-BSA 0.2%-Triton for 5 min. Before and after hybridization with the primary antibody (anti-VE-cadherin Ab-33168 “Abcam” - Cambridge, UK) cells were washed with PBS 1%-BSA. Secondary antibody hybridization was performed using Alexa Fluor® 488 labeled anti-rabbit (Thermo Scientific, Waltham, MA, USA). Nuclei were stained using DAPI (Figs 3A and 4C). Images were taken using an Inverted Nikon FLUO-Scope (Nikon, Tokyo, Japan). Data were analyzed with Nikon software ND2. For particle immunofluorescence, samples were prepared as described above for flow cytometry. After particle conjugation with antibody, 105 particles were seeded on an 8-well Nunc® Lab-Tek® Chamber Slide™ (Thermo Scientific). Images were acquired with a Nikon A1 confocal imaging system and analyzed with Nikon NIS Elements software (Nikon).
Palomba R., Parodi A., Evangelopoulos M., Acciardo S., Corbo C., de Rosa E., Yazdi I.K., Scaria S., Molinaro R., Furman N.E., You J., Ferrari M., Salvatore F, & Tasciotti E. (2016). Biomimetic carriers mimicking leukocyte plasma membrane to increase tumor vasculature permeability. Scientific Reports, 6, 34422.
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4

Comprehensive Immunostaining Protocol for Endothelial Markers

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The anti-Trio (H-120, 1:500), anti-Notch1 ICD (C-20-R, 1:1000), anti-Notch1 ECD (H-131, 1:500), anti-VE-cadherin (F-8, 1:200), and anti-VE-PTP (H-300, 1:250) antibodies were from Santa Cruz Biotechnology. Anti-VE-cadherin (ab33168, 1:2,000), anti-RFP (ab124754, 1:1000), and anti-β tubulin (ab6046, 1:5,000) antibodies were from Abcam. Anti-VEGFR2 antibody (55B11, 1:1000), anti-Notch1 V1754G (D3B8, 1:1000 WB), anti-HA (6E2, 1:1000 WB, AF647 conjugate IF) was from Cell Signaling Technologies. Anti-Rac1 antibody (102, 1:1,000) was from BD Biosciences. Rhodamine and Alexa Fluor 488-labelled phalloidin, Alexa Fluor-488, 568 and 647 goat anti-mouse and anti-rabbit IgG secondary antibodies were from Life Technologies. HRP-conjugated donkey anti-mouse, anti-rabbit, and anti-goat IgG secondary antibodies were from Fitzgerald. Anti-PTPRF antibody (SAB4200321, 1:1000), DAPI, DAPT, Evans Blue dye, dynasore hydrate, and formamide were from Sigma. Rac1 inhibitor NSC 23766 was from Santa Cruz Biotechnology.
Polacheck W.J., Kutys M.L., Yang J., Eyckmans J., Wu Y., Vasavada H., Hirschi K.K, & Chen C.S. (2017). A non-canonical Notch complex regulates adherens junctions and vascular barrier function. Nature, 552(7684), 258-262.
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5

Comprehensive Immunostaining Protocol for Endothelial Markers

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The anti-Trio (H-120, 1:500), anti-Notch1 ICD (C-20-R, 1:1000), anti-Notch1 ECD (H-131, 1:500), anti-VE-cadherin (F-8, 1:200), and anti-VE-PTP (H-300, 1:250) antibodies were from Santa Cruz Biotechnology. Anti-VE-cadherin (ab33168, 1:2,000), anti-RFP (ab124754, 1:1000), and anti-β tubulin (ab6046, 1:5,000) antibodies were from Abcam. Anti-VEGFR2 antibody (55B11, 1:1000), anti-Notch1 V1754G (D3B8, 1:1000 WB), anti-HA (6E2, 1:1000 WB, AF647 conjugate IF) was from Cell Signaling Technologies. Anti-Rac1 antibody (102, 1:1,000) was from BD Biosciences. Rhodamine and Alexa Fluor 488-labelled phalloidin, Alexa Fluor-488, 568 and 647 goat anti-mouse and anti-rabbit IgG secondary antibodies were from Life Technologies. HRP-conjugated donkey anti-mouse, anti-rabbit, and anti-goat IgG secondary antibodies were from Fitzgerald. Anti-PTPRF antibody (SAB4200321, 1:1000), DAPI, DAPT, Evans Blue dye, dynasore hydrate, and formamide were from Sigma. Rac1 inhibitor NSC 23766 was from Santa Cruz Biotechnology.
Polacheck W.J., Kutys M.L., Yang J., Eyckmans J., Wu Y., Vasavada H., Hirschi K.K, & Chen C.S. (2017). A non-canonical Notch complex regulates adherens junctions and vascular barrier function. Nature, 552(7684), 258-262.
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