Treatment of cells with TNFα (Life Technologies, Frederick, MD) consisted of overnight incubation at 20 ng/ml. S1P (Sigma-Aldrich, St. Louis, MO) treatment was at 0.5 μM for 5 min. Anti–ICAM-1 antibodies included mouse BBIG-I1 from R&D Systems (Minneapolis, MN) and rabbit H-108 from Santa Cruz Biotechnology (Dallas, TX). Antibodies to VE-cadherin included a monoclonal mouse anti–human CD144 antibody (clone 55-7H1; BD Biosciences, San Jose, CA) and a polyclonal rabbit anti–human VE-cadherin antibody (#2158; Cell Signaling, Danvers, MA). Anti-GFP was a rabbit polyclonal (ab6556; Abcam, Cambridge, MA). Anti-Arp3 was either a mouse anti–human monoclonal (FMS338; Sigma-Aldrich, St. Louis, MO) or a rabbit anti–bovine polyclonal (Welch et al., 1997 (link))). Rabbit anti–human WAVE-2 monoclonal antibody (D2C8, #3659) was from Cell Signaling. Mouse anti–human ZO-1 monoclonal antibody (ZO1-1A12) was from Invitrogen (Camarillo, CA). Mouse anti–human glyceraldehyde-3-phosphate dehydrogenase monoclonal antibody (6C5) was from Abcam. Mouse anti–human vinculin monoclonal antibody (hVIN-1) was from Sigma-Aldrich. Rabbit anti–nonmuscle myosin heavy chain II-A polyclonal antibody (PRB-440P) was from Covance (Emeryville, CA). Secondary antibodies and phalloidin, fluorescently conjugated with Alexa dyes, were obtained from Molecular Probes (Life Technologies, Eugene, OR).
Anti human ve cadherin antibody
Manufactured by Cell Signaling Technology
The Anti–human VE-cadherin antibody is a laboratory tool used to detect and study the VE-cadherin protein. VE-cadherin is an adhesion molecule found in the cell-cell junctions of endothelial cells. This antibody can be used to identify and quantify VE-cadherin expression in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Sphingosine 1 phosphate (s1p), by Merck Group (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Ab6556, by Abcam (1 mentions)
Axio observer z1, by Zeiss (1 mentions)
Axiovision software, by Zeiss (1 mentions)
Af623, by R&D Systems (1 mentions)
Rhodamine conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
2 protocols using anti human ve cadherin antibody
1
Immunofluorescence Staining for Endothelial Cell Junctions
2
Endothelial Cell Barrier Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human brain microvascular endothelial cells (HBMECs) were seeded onto fibronectin pre-coated 12-well μ-slides (ibidi GmbH, Martinsried, Germany) with a cell density of 8 × 104/mL for overnight culturing. HBMECs were then treated with concentrated conditioned medium from control and knockdown cells for 24 h. To neutralize ANGPT2, the anti-ANGPT2 antibody (10 μg/mL; AF623, R&D) was added to the concentrated conditioned medium for 1 h of incubation, and then this medium was used to treat HBMECs. After culturing, cells were fixed with 4% paraformaldehyde in PBS for 20 min at room temperature and permeabilized with 0.1% Triton X-100 in PBS (PBS-T) at 4 °C for 10 min. After blocking with 4% BSA in PBS at 37 °C for 30 min, cells were stained with anti-human VE-cadherin antibody (#2158, Cell Signaling) and rhodamine conjugated anti-rabbit IgG (Jackson ImmunoResearch Laboratories, West Grove, PA). DAPI was used for nucleus staining. Slides were mounted and viewed under a fluorescence microscope (Zeiss Axio Observer Z1) and analyzed using the AxioVision software (Zeiss). The length of VE-cadherin reflected by fluorescence was calculated by the ImageJ program and normalized to the cell number.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!