All lanes : Anti-Neurofilament antibody [RNF406] (ab134306) at 1/100 dilution

Lane 1 : Wild-type HEK293 whole cell lysate
Lane 2 : Neurofilament knockout HEK293 whole cell lysate
Lane 3 : A549 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 102 kDa
Observed band size: 140-160 kDa why is the actual band size different from the predicted?

Lanes 1 - 3: Merged signal (red and green). Green - ab134306 observed at 140-160 kDa kDa. Red - loading control, ab52866, observed at 50 kDa.

ab134306 was shown to specifically react with Neurofilament in wild-type HEK293 cells as signal was lost in Neurofilament knockout cells. Wild-type and Neurofilament knockout samples were subjected to SDS-PAGE. ab134306 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1/100 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human brain cortex tissue labelling 140 kD + 160 kD Neurofilament with ab134306 at 10 µg/ml, using a biotinylated horse anti-goat IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.