All lanes : Anti-TEF1/TEAD-1 antibody [EPR3968(2)] (ab133535) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : TEAD1 knockout A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 48 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Lanes 1 - 2: Merged signal (red and green). Green - ab133535 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab133535 was shown to react with TEF1/TEAD-1 in wild-type A549 cells in western blot with loss of signal observed in TEAD1 knockout sample. Wild-type and TEAD1 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab133535 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 incubated 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.

Immunofluorescence analysis of HeLa cells labelling TEF1/TEAD-1 with ab133535 at 1/50

All lanes : Anti-TEF1/TEAD-1 antibody [EPR3968(2)] (ab133535) at 1/1000 dilution

Lane 1 : HeLa cell lysates
Lane 2 : A431 cell lysates
Lane 3 : 293T cell lysates
Lane 4 : Fetal muscle lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 48 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Equilibrium disassociation constant (K_D)
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