Anti-VEGFA 抗体

• WB

Project3318****

Western blot - Anti-VEGFA antibody (AB46154)

All lanes:

Western blot - Anti-VEGFA antibody (ab46154) at 0.5 µg/mL

Lanes 1 and 8:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lanes 2 and 9:

Western blot - Jurkat whole cell lysate (<a href='/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 10 µg

Lanes 3 and 10:

Western blot - A-431 whole cell lysate (<a href='/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 10 µg

Lanes 4 and 11:

Western blot - HEK-293 whole cell lysate (<a href='/products/cell-lysates/hek-293-whole-cell-lysate-ab7902'>ab7902</a>) at 10 µg

Lanes 5 and 12:

Hep G2 whole cell lysate (ab7900) at 10 µg

Lanes 6 and 13:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lanes 7 and 14:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

false

• WB

Lab

Western blot - Anti-VEGFA antibody (AB46154)

This blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab46154 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-VEGFA antibody (ab46154) at 1 µg/mL

All lanes:

HCT116 whole cell lysate at 20 µg

Secondary

All lanes:

Rabbit IgG secondary antibody at 1/10000 dilution

Predicted band size: 27 kDa

Observed band size: 23 kDa

true

Exposure time: 2min

• ELISA

Lab

ELISA - Anti-VEGFA antibody (AB46154)

ab46154 was tested using indirect ELISA. The wells were coated with peptide (1 μg x mL^-1 at 100 μL per well) overnight at 4°C, followed by a 1% fat-free milk blocking step for 1 hour at room temperature. The primary antibody (ab46154) was added at a range of dilutions (50 μL per well) for 1 hour at room temperature. ab97080 (Goat Anti-Rabbit IgG H&L-HRP) was used as a secondary antibody at 1 : 50.000 dilution for 1 hour at room temperature and signal was developed with TMB substrate.

• WB

Lab

Western blot - Anti-VEGFA antibody (AB46154)

This blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab46154 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-VEGFA antibody (ab46154) at 1 µg/mL

Lane 1:

Recombinant Human VEGFA protein (His tag) (ab204773) at 0.1 µg

Lane 2:

Western blot - Recombinant mouse VEGFA protein (Active) (<a href='/products/proteins-peptides/recombinant-mouse-vegfa-protein-active-ab185265'>ab185265</a>) at 0.1 µg

Secondary

All lanes:

Rabbit IgG secondary antibody at 1/10000 dilution

Predicted band size: 27 kDa

Observed band size: 23 kDa

true

Exposure time: 10s