重组Anti-VE Cadherin抗体[RM2022] (ab318152)

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

Negative control: Hela (PMID: 35164755), PANC-1 

The molecular weight observed is consistent with what has been described in the literature (PMID: 36631513).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade  (ab176842) staining at 1/100000 dilution.

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

Samples are non-boiled as boiling may cause protein aggregation.

Exposure time: Lane 1: 3 seconds, Lanes 2-3: 6 seconds

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

Samples are non-boiled as boiling may cause protein aggregation.

Exposure time: Lanes 1-2 6 seconds, Lane 3-4: 15 seconds

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

Negative control: C2C12

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade  (ab176842) staining at 1/100000 dilution.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Membranous staining on endothelium of human kidney (PMID: 29846473). The section was incubated with ab318152 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Membranous staining on endothelium of human cardiac muscle. The section was incubated with ab318152 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Membranous staining on endothelium of human breast cancer (PMID: 18316602). The section was incubated with ab318152 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Membranous staining on endothelium of mouse lung. The section was incubated with ab318152 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Membranous staining on endothelium of mouse cardiac muscle. The section was incubated with ab318152 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond ™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in HUVEC cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control: Hela (PMID: 35164755).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor ® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed at 1/1000 2 ug/ml dilution.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized bEnd.3 (mouse brain endothelial cell) cells labelling VE Cadherin with ab318152 at 1/500 (1.028 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in bEnd.3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control: C2C12
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor ® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed at 1/1000 2 ug/ml dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (mouse myoblast, Left) / bEnd.3 (mouse brain endothelial cell, Right) cells labelling VE Cadherin with ab318152 at 1/500 dilution (0.1 ug)/Magenta compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor ® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control: C2C12.

VE Cadherin was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab318152 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318152 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 2: ab318152 IP in HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab318152 in HUVEC whole cell lysate

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

Exposure time: 3 seconds.