Anti-Vinculin antibody [EPR19579] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR19579] - BSA and Azide free (AB236031)

Blocking/Dilution buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207440).

All lanes:

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (<a href='/en-us/products/primary-antibodies/vinculin-antibody-epr19579-loading-control-ab207440'>ab207440</a>) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 124 kDa

Observed band size: 124 kDa

false

Exposure time: 30s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR19579] - BSA and Azide free (AB236031)

This data was developed using the same antibody clone in a different buffer formulation (ab207440).

Lanes 1 - 4 : Merged signal (red and green). Green - ab207440 observed at 124 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab207440 was shown to react with VCL in A431 wild-type cells in Western blot. Loss of signal was observed when VCL knockout sample was used. A431 wild-type and VCL knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab207440 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (<a href='/en-us/products/primary-antibodies/vinculin-antibody-epr19579-loading-control-ab207440'>ab207440</a>) at 1/1000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

VCL knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human VCL (Vinculin) knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-vcl-vinculin-knockout-a-431-cell-line-ab261898'>ab261898</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Predicted band size: 124 kDa

Observed band size: 124 kDa

false