All lanes : Anti-Transferrin Receptor antibody [EPR4012] (ab108985) at 1/1000 dilution

Lane 1 : Human placenta cell lysate
Lane 2 : TF-1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : A549 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1 & 4 : HRP labelled goat anti-rabbit at 1/2000 dilution


Predicted band size: 84 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

Overlay histogram showing Jurkat cells stained with ab108985 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108985, 1/100) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.