All lanes : Anti-APH1a antibody ( ab12104 ) at 1/500 dilution Lane 1 : 20ug human brain lysate Lane 2 : 20ug mouse brain lysate Lane 3 : 20ug human brain lysate with Human APH1a peptide (ab12394) at 1 µg/ml Lane 4 : 20ug mouse brain lysate with Human APH1a peptide (ab12394) at 1 µg/ml Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ( ab7090 ) at 1/5000 dilution Performed under reducing conditions. Predicted band size : 30 kDa Observed band size : 30 kDa Additional bands at : 31 kDa,75 kDa. We are unsure as to the identity of these extra bands. Exposure time : 30 seconds Rabbit polyclonal to APH1a ( ab12104 ) used at 1/500 on: Lane 1: Human Brain Lysate Lane 2: Mouse Brain Lysate Lane 3: Human Brain Lysate + 1µg/ml APH1a blocking peptide (ab12394) Lane 4: Mouse Brain Lysate + 1µg/ml APH1a blocking peptide (ab12394) Secondary antibody : Goat polyclonal to Rabbit IgG (HRP) - pre-adsorbed at 1/5000 ( ab7090 ). Expected molecular weight was 30kDa. The band observed between 20-25kDa is
All lanes : Anti-APH1a antibody ( ab12104 ) at 1 µg/ml Lane 1 : Mouse brain lysate Lane 2 : Rat brain lysate Lane 3 : Mouse brain lysate with Human APH1a peptide (ab12394) at 1 µg/ml Lane 4 : Rat brain lysate with Human APH1a peptide (ab12394) at 1 µg/ml Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ( ab7090 ) at 1/5000 dilution Performed under reducing conditions. Observed band size : 37 kDa ( why is the actual band size different from the predicted? ) Additional bands at : 21 kDa,75 kDa. We are unsure as to the identity of these extra bands. Exposure time : 1 minute