Anti-VEGF Receptor 2 (phospho Y1214) 抗体

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 2 (phospho Y1214) antibody (AB5475)

Modulation of expression of target protein by cell treatment to demonstrate antibody specificity.

The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate

All lanes:

Western blot - Anti-VEGF Receptor 2 (phospho Y1214) antibody (ab5475) at 1/1000 dilution

Lane 1:

HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 2:

HT-29 Serum Starved followed by treatment for 10 minutes with 100 ng/mL of SCF at 30 µg

Lane 3:

Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 4:

Caco-2 Serum Starved followed by treatment for 10 minutes with 100 ng/mL of SCF at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/5000 dilution

Predicted band size: 151 kDa

Observed band size: ~151 kDa

false

• WB

Unknown

Western blot - Anti-VEGF Receptor 2 (phospho Y1214) antibody (AB5475)

Peptide Competition and Stimulation : Extracts prepared from CSF-1 stimulated (1-4, 6) or unstimulated (5) PAE cells transfected with a chimeric CSF-1/VEGFR2 receptor were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 μg/mL ab5475 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with : no peptide (1, 5, 6), the nonphosphopeptide corresponding to the immunogen (2), a generic phosphotyrosine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab5475 blocks the antibody signal, and the stimulation of the phospho signal after stimulating ligand is added (CSF-1 for the chimeric r

All lanes:

Western blot - Anti-VEGF Receptor 2 (phospho Y1214) antibody (ab5475)

Predicted band size: 151 kDa

false