Cell-laden hollow filaments were fixed with ice-cold methanol for 10 min at −20 °C. The constructs were halved lengthwise with a razor blade, washed three times for 5 min with washing buffer (0.5% Tween 20 in PBS*) and blocked with 3% goat serum (Jackson ImmunoResearch Europe Ltd) in washing buffer for 30 min. Rabbit anti-VE cadherin antibody (Cell Signaling Technologies, catalog #2500S) was diluted 1:100 in blocking buffer (3% normal goat serum in washing buffer) and incubated on the samples overnight at 4 °C. The constructs were washed three times for 5 min with washing buffer and goat-anti rabbit IgG-FITC (Santa Cruz, 1:200 in washing buffer) was added for 1 h at RT. Finally, samples were washed again three times for 5 min in washing buffer, stained with DAPI (Thermo Fisher Scientific, 1:1000 in washing buffer) and analyzed after another washing step using a cLSM (Zeiss, LSM800; magnification 400-fold).
Rabbit anti ve cadherin antibody
Manufactured by Cell Signaling Technology
Sourced in Japan
Rabbit anti-VE cadherin antibody is a primary antibody that specifically binds to and recognizes the vascular endothelial cadherin (VE-cadherin) protein. VE-cadherin is a cell-cell adhesion molecule that plays a critical role in the organization and function of vascular endothelial cells.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 800, by Zeiss (1 mentions)
Goat anti rabbit igg fitc, by Santa Cruz Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Normal goat serum (ngs), by Cell Signaling Technology (1 mentions)
Goat serum, by Jackson ImmunoResearch (1 mentions)
Triton x 100, by Merck Group (1 mentions)
24 well culture plate, by Corning (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Fujifilm (1 mentions)
Alexa fluor 647 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Pi double staining kit, by Dojindo Laboratories (1 mentions)
2 protocols using rabbit anti ve cadherin antibody
1
Immunofluorescent Staining of Cell-Laden Filaments
2
Visualizing VE-cadherin in HUVEC Monolayers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To
investigate the effects of [Fe(CN)6]4– on HUVEC monolayers, VE-cadherin in the HUVEC monolayers was stained.
First, HUVECs were cultured in a 24-well culture plate (Corning) for
7 days to prepare HUVEC monolayers. The monolayers were washed with
PBS three times and then immersed in PBS containing 4% paraformaldehyde
(Fujifilm Wako Pure Chemical Co., Japan) for 15 min. Next, after washing
with PBS three times, the monolayers were immersed in 0.1% Triton
X-100 (Sigma-Aldrich Japan, Japan) for 15 min. After removing Triton
X-100, the monolayers were immersed in PBS containing 1% bovine serum
albumin (Sigma-Aldrich Japan) and rabbit anti VE-cadherin antibody
(1:500 dilution, Cell Signaling Technology, USA). After incubating
overnight at 4 °C, the solution was replaced with PBS containing
a secondary antibody, Alexa Fluor 647 conjugated goat anti-rabbit
IgG (1:1000 dilution, Thermo Fisher Scientific, USA), and 45 μM
propidium iodide (PI, double staining kit, Dojindo, Japan). The monolayers
were incubated overnight at 4 °C and then observed under a fluorescence
microscope (Leica DMi8, Leica, Germany).
investigate the effects of [Fe(CN)6]4– on HUVEC monolayers, VE-cadherin in the HUVEC monolayers was stained.
First, HUVECs were cultured in a 24-well culture plate (Corning) for
7 days to prepare HUVEC monolayers. The monolayers were washed with
PBS three times and then immersed in PBS containing 4% paraformaldehyde
(Fujifilm Wako Pure Chemical Co., Japan) for 15 min. Next, after washing
with PBS three times, the monolayers were immersed in 0.1% Triton
X-100 (Sigma-Aldrich Japan, Japan) for 15 min. After removing Triton
X-100, the monolayers were immersed in PBS containing 1% bovine serum
albumin (Sigma-Aldrich Japan) and rabbit anti VE-cadherin antibody
(1:500 dilution, Cell Signaling Technology, USA). After incubating
overnight at 4 °C, the solution was replaced with PBS containing
a secondary antibody, Alexa Fluor 647 conjugated goat anti-rabbit
IgG (1:1000 dilution, Thermo Fisher Scientific, USA), and 45 μM
propidium iodide (PI, double staining kit, Dojindo, Japan). The monolayers
were incubated overnight at 4 °C and then observed under a fluorescence
microscope (Leica DMi8, Leica, Germany).
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