Anti-VEGF Receptor 2/Kdr Antibody Picoband®

Figure 1. Western blot analysis of VEGF Receptor 2 using anti-VEGF Receptor 2 antibody (A00901-3).
The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Marker
Lane 2: mouse heart tissue lysates,
Lane 3: mouse thymus tissue lysates,
Lane 4: mouse spleen tissue lysates,
Lane 5: mouse kidney tissue lysates.
Lane 6: mouse brain tissue lysates,
Lane 5: mouse lung tissue lysates.
Lane 7: mouse HEPA1-6 whole cell lysates,
Lane 8: mouse NIH3T3 whole cell lysates.
The membrane was incubated with rabbit anti-VEGF Receptor 2 antigen affinity purified polyclonal antibody (Catalog # A00901-3) at 1.0 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system.

Figure 2. IHC analysis of VEGF Receptor 2 using anti-VEGF Receptor 2 antibody (A00901-3).
VEGF Receptor 2 was detected in paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-VEGF Receptor 2 Antibody (A00901-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of VEGF Receptor 2 using anti-VEGF Receptor 2 antibody (A00901-3).
VEGF Receptor 2 was detected in paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-VEGF Receptor 2 Antibody (A00901-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 4. IF analysis of VEGF Receptor 2 using anti-VEGF Receptor 2 antibody (A00901-3).
VEGF Receptor 2 was detected in immunocytochemical section of NIH3T3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-VEGF Receptor 2 Antibody (A00901-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 5. Flow Cytometry analysis of MFC cells using anti-VEGF Receptor 2 antibody (A00901-3).
Overlay histogram showing MFC cells stained with A00901-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-VEGF Receptor 2 Antibody (A00901-3,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 6. Flow Cytometry analysis of LLC cells using anti-VEGF Receptor 2 antibody (A00901-3).
Overlay histogram showing LLC cells stained with A00901-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-VEGF Receptor 2 Antibody (A00901-3,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.