Anti-Vinculin antibody [EPR19579] - Loading Control

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (AB207440)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (ab207440) 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] - Loading Control (AB207440)

Lanes 1-4 : Merged signal (red and green). Green - ab207440 observed at 124 kDa. Red - loading control ab7291 observed at 50 kDa.

ab207440 Anti-Vinculin antibody [EPR19579] was shown to specifically react with Vinculin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265580 (knockout cell lysate ab257795) was used. Wild-type and Vinculin knockout samples were subjected to SDS-PAGE. ab207440 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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 (ab207440) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Vinculin knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human VCL (Vinculin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-vcl-vinculin-knockout-hela-cell-line-ab265580'>ab265580</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 124 kDa

Observed band size: 124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (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 (ab207440) 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

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (AB207440)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-4 : 3 minutes; Lane 5/6 : 5 seconds.

All lanes:

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (ab207440) at 1/2000 dilution

Lane 1:

K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 3:

HUVEC (Human umbilical vein endothelial cell line) whole cell lysate at 10 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 5:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 6:

Human testis lysate at 10 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 1/100000 dilution

Predicted band size: 124 kDa

Observed band size: 124 kDa

false

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (AB207440)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1-3 : 30 seconds; Lane 4-8 : 10 seconds.

All lanes:

Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (ab207440) at 1/1000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat kidney lysate at 10 µg

Lane 5:

Rat spleen lysate at 10 µg

Lane 6:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 7:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 8:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 124 kDa

Observed band size: 124 kDa

false