Synthetic peptide corresponding to Human VE Cadherin aa 750 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available as
Our Abpromise guarantee covers the use of ab33168 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 - 5 µg/ml.
Abcam recommends using this product with confluent cells.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 115 kDa (predicted molecular weight: 87 kDa).Can be blocked with VE Cadherin peptide (ab27462).
Abcam recommends using BSA blocking with this product. Milk blocking will give a greatly reduced signal strength in WB.
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|In-Cell ELISA||Use at an assay dependent concentration. PubMed: 22689949|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
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.
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.
ab33168 stained HUVEC cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour 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) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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).