Anti-PDK1 antibody [EPR13009] - BSA and Azide free

• WB

Lab

Western blot - Anti-PDK1 antibody [EPR13009] - BSA and Azide free (AB249846)

This data was developed using the same antibody clone in a different buffer formulation (ab175190).

Lanes 1- 2 : Merged signal (red and green). Green - ab175190 observed at 46 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab175190 was shown to react with PDK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264791 (knockout cell lysate ab257164) was used. Wild-type HeLa and PDK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab175190 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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) 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-PDK1 antibody [EPR13009] (<a href='/en-us/products/primary-antibodies/pdk1-antibody-epr13009-ab175190'>ab175190</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PDK1 knockout HeLa cell lysate at 20 µg

Predicted band size: 49 kDa

Observed band size: 46 kDa

false

• WB

Lab

Western blot - Anti-PDK1 antibody [EPR13009] - BSA and Azide free (AB249846)

This data was developed using the same antibody clone in a different buffer formulation (ab175190).

Lanes 1 - 4 : Merged signal (red and green). Green - ab175190 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab175190 was shown to specifically recognize PDK1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when PDK1 knockout samples were examined. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. ab175190 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PDK1 antibody [EPR13009] - BSA and Azide free (ab249846) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

PDK1 knockout HAP1 cell lysate at 20 µg

Lane 3:

Human fetal heart lysate at 20 µg

Lane 4:

LnCaP cell lysate at 20 µg

Predicted band size: 49 kDa

false