Anti-VE Cadherin antibody - Intercellular Junction Marker (ab33168)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab33168 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

The band we observe at 115 kDa is believed to be the glycosylated form of the protein.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab33168 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

The band we observe at 115 kDa is believed to be the glycosylated form of the protein.

ab33168 stained in HUVEC cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab33168 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

This image was generated using confluent cells.

ab33168 used in Flow cytometry.
Human REH B cells were fixed in paraformaldehyde and permeabilized using saponin. Primary antibody used undiluted (2µl in 100µl of cells in PBS) and incubated for 15 minutes at 4°C. The secondary antibody used was an undiluted, Alexa Fluor®488 conjugated goat anti-rabbit IgG.

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ICC/IF image of VE Cadherin stained HUVEC cells. The cells were incubated with the antibody ab33168 at 1/150 (Green). The cells were also stained with Rhodamine phalloidin (Red).

The observed band for Cadherin 5 has a higher molecular weight of 115kDa due to glycosylation of the protein. 

The immunogen used to raise this antibody has 89% homology with Cadherin 18, 88kDa , which we believe is the additional observed band at 117kDa, again due to glycosylation of the protein. 

ab33168 Immunoprecipitating VE Cadherin in human HUVEC whole cell lysate. 1000000 cells lysate was incubated with primary antibody (1/100 in 0.5% NP40, 150mM NaCl, 50mM Tris) and matrix (Dynabeads) for 2 hours at 4°C. For western blotting a HRP-conjugated mouse anti-VE Cadherin (1/3000) was used to confirm successful immunoprecipation.

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