Anti-VEGF Receptor 2 antibody [EPR28328-80]

• IP

Supplier Data

Immunoprecipitation - Anti-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

VEGF Receptor 2 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab315238 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315238 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 : ab315238 IP in HUVEC (human umbilical vein endothelial cell) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315238 in HUVEC whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-VEGF Receptor 2 antibody [EPR28328-80] (ab315238) 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 1/5000 dilution

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

Immunohistochemical analysis of paraffin-embedded Mouse placenta tissue labeling VEGF Receptor 2 with ab315238 at 1/1000 (0.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse placenta. The section was incubated with ab315238 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-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling VEGF Receptor 2 with ab315238 at 1/1000 (0.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse kidney. The section was incubated with ab315238 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-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling VEGF Receptor 2 with ab315238 at 1/1000 (0.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of rat liver. The section was incubated with ab315238 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-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling VEGF Receptor 2 with ab315238 at 1/1000 (0.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of mouse liver. The section was incubated with ab315238 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-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling VEGF Receptor 2 with ab315238 at 1/1000 (0.505 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelial cells of rat kidney. The section was incubated with ab315238 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

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

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

Exposure time : Lane 1 : 10 seconds; Lanes 2-3 : 180 seconds.

All lanes:

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (ab315238) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

EA.hy926 (human somatic cell hybrid endothelial cell) whole cell 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: 230 kDa,200 kDa

false

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

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

Negative control : HeLa, A549 (PMID : 22514712), K-562 (PMID : 10953026).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (ab315238) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

A549 (human lung carcinoma 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 1/20000 dilution

Observed band size: 230 kDa,200 kDa,124 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

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

All lanes:

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (ab315238) at 1/1000 dilution

Lane 1:

Mouse placenta tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse lung tissue lysate at 20 µg

Lane 5:

Rat placenta tissue lysate at 20 µg

Lane 6:

Rat spleen tissue lysate at 20 µg

Lane 7:

Rat liver tissue 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 1/20000 dilution

Observed band size: 230 kDa,200 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (AB315238)

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

This antibody does not cross-react with human VEGF Receptor 1 & VEGF Receptor 3.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-VEGF Receptor 2 antibody [EPR28328-80] (ab315238) at 1/1000 dilution

Lane 1:

His-tagged human VEGF Receptor 1 fragment at 10 ng

Lane 2:

His-tagged human VEGF Receptor 2 fragment at 10 ng

Lane 3:

His-tagged human VEGF Receptor 3 fragment at 10 ng

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 1/20000 dilution

Observed band size: 30 kDa

false

Exposure time: 10s