Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling VE Cadherin with ab313632 at 1/50 (10.32 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.Negative control : HeLa (PMID : 35164755).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling VE Cadherin with ab313632 at 1/2000 (0.258 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on human lung (PMID : 29846473). The section was incubated with ab313632 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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation. Flow cytometric analysis of HeLa (human cervical adenocarcinoma epithelial cell, Left) / HUVEC (human umbilical vein endothelial cell, Right) cells labelling VE Cadherin with ab313632 at 1/500 dilution (0.1 ug)/Red 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 : HeLa (PMID : 35164755).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VE Cadherin with ab313632 at 1/2000 (0.258 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on vascular endothelium of human kidney (PMID : 29846473). The section was incubated with ab313632 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling VE Cadherin with ab313632 at 1/2000 (0.258 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on vascular endothelium of human colon cancer (PMID : 27466381). The section was incubated with ab313632 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

• IP

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Immunoprecipitation - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation. VE Cadherin was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab313632 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313632 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 : ab313632 IP in HUVEC (human umbilical vein endothelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313632 in HUVEC whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds The molecular weight observed is consistent with what has been described in the literature (PMID : 36631513).

All lanes:

Immunoprecipitation - Anti-VE Cadherin antibody [EPR27436-55] (<a href='/en-us/products/primary-antibodies/ve-cadherin-antibody-epr27436-55-ab313632'>ab313632</a>) at 1/30 dilution

All lanes:

HUVEC (human umbilical vein endothelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

false

Exposure time: 3s

• WB

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Western blot - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation.

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

Negative control : Hela (PMID : 35164755).

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

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

In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution.

All lanes:

Western blot - Anti-VE Cadherin antibody [EPR27436-55] (<a href='/en-us/products/primary-antibodies/ve-cadherin-antibody-epr27436-55-ab313632'>ab313632</a>) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) 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 1/100000 dilution

Observed band size: 87-100 kDa,130 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

This data was developed using ab313632, the same antibody clone in a different buffer formulation.

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

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

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

All lanes:

Western blot - Anti-VE Cadherin antibody [EPR27436-55] (<a href='/en-us/products/primary-antibodies/ve-cadherin-antibody-epr27436-55-ab313632'>ab313632</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 87-100 kDa,130 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-VE Cadherin antibody [EPR27436-55] - BSA and Azide free (AB313633)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-VE Cadherin antibody [EPR27436-55] (ab313633) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-VE Cadherin (phospho Y685) antibody [EPR27437-539] (<a href='/en-us/products/primary-antibodies/ve-cadherin-phospho-y685-antibody-epr27437-539-ab325018'>ab325018</a>) at 1/1000 dilution

Lane 1:

Untreated HUVEC (human umbilical vein endothelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HUVEC treated with 100 μM Pervanadate for 10 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated HUVEC whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

HUVEC treated with 100 μM Pervanadate for 10 minutes whole cell lysate (alkaline phosphatase treated membrane) 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 1/100000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 26s