Anti-VASP antibody [EPR1337(2)] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-VASP antibody [EPR1337(2)] - BSA and Azide free (AB231823)

ab109321 (purified) at 1/30 immunoprecipitating VASP in HEK293 cell lysate (Lane 1). Lane 2 - rabbit monoclonal IgG instead of ab109321 in HEK293 lysates. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109321).

All lanes:

Immunoprecipitation - Anti-VASP antibody [EPR1337(2)] (<a href='/en-us/products/primary-antibodies/vasp-antibody-epr13372-ab109321'>ab109321</a>)

Predicted band size: 39 kDa

Observed band size: 46 kDa

false

• WB

Unknown

Western blot - Anti-VASP antibody [EPR1337(2)] - BSA and Azide free (AB231823)

This data was developed using ab109321, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab109321 observed at 46 kDa. Red - loading control ab8245 observed at 36 kDa.

ab109321 Anti-VASP antibody [EPR1337(2)] was shown to specifically react with VASP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265892 (knockout cell lysate ab257792) was used. Wild-type and VASP knockout samples were subjected to SDS-PAGE. ab109321 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-VASP antibody [EPR1337(2)] (<a href='/en-us/products/primary-antibodies/vasp-antibody-epr13372-ab109321'>ab109321</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 2:

Western blot - Human VASP knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-vasp-knockout-hela-cell-line-ab265892'>ab265892</a>)

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole 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 1/10000 dilution

Predicted band size: 39 kDa

Observed band size: 46 kDa

false