Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (AB205336)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized bEnd.3 (Mouse brain microvascular endothelial cell line) cells labeling VE Cadherin with ab205336 at 1/1000 dilution, followed by Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing membrane staining on bEnd.3 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody -Loading control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (AlexaFluor®594 ) preadsorbed (ab150120) at 1/500 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab205336 at 1/1000 dilution followed by ab150120 at 1/500 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/500 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (AB205336)

VE Cadherin was immunoprecipitated from 1mg of bEnd.3 (Mouse brain microvascular endothelial cell line) whole cell lysate with ab205336 at 1/80 dilution.

Western blot was performed from the immunoprecipitate using ab205336 at 1/1000 dilution.

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

Lane 1 : bEnd.3 whole cell lysate 10ug (Input).

Lane 2 : ab205336 IP in bEnd.3 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab205336 in bEnd.3 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (ab205336)

Predicted band size: 88 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (AB205336)

VE Cadherin was immunoprecipitated from 1mg of Mouse lung whole cell lysate with ab205336 at 1/80 dilution.

Western blot was performed from the immunoprecipitate using ab205336 at 1/1000 dilution.

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

Lane 1 : Mouse lung whole cell lysate 10ug (Input).

Lane 2 : ab205336 IP in Mouse lung whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab205336 in Mouse lung whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

Due to a high degree of glycosylation and phosphorylation, the observed MW is higher than the predicted MW. The 90kDa fragment represents the extracellular domain where the immunogen is located.

All lanes:

Immunoprecipitation - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (ab205336)

Predicted band size: 88 kDa

false

• WB

Supplier Data

Western blot - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (AB205336)

Blocking/Dilution buffer : 5% NFDM/TBST.

Due to a high degree of glycosylation and phosphorylation, the observed MW is higher than the predicted MW. The 90kDa fragment represents the extracellular domain where the immunogen is located.

All lanes:

Western blot - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (ab205336) at 1/1000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 88 kDa

Observed band size: 120 kDa,90 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (AB205336)

Blocking/Dilution buffer : 5% NFDM/TBST.

Due to a high degree of glycosylation and phosphorylation, the observed MW is higher than the predicted MW. The 90kDa fragment represents the extracellular domain where the immunogen is located.

All lanes:

Western blot - Anti-VE Cadherin antibody [EPR18229] - Intercellular Junction Marker (ab205336) at 1/1000 dilution

Lane 1:

Mouse lung lysate at 20 µg

Lane 2:

Mouse placenta lysate at 20 µg

Lane 3:

bEnd.3 (Mouse brain microvascular endothelial cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 88 kDa

Observed band size: 125 kDa,90 kDa

false

Exposure time: 1s