Comparison of nitrocellulose and PVDF membranes in immunoblotting of α-syn.

<p>Immunoblotting was performed using a PVDF membrane (left panels) and a nitrocellulose membrane (right panels). Briefly, total cell lysates were prepared from SK-MEL28 (positive control), SH-SY5Y (positive control), and BJAB (negative control). The protein samples (∼10 µg) were loaded onto a 15% SDS-polyacrylamide gel and electrophoresed, followed by Western transfer onto a PVDF or nitrocellulose membrane. The membrane was then fixed with or without 0.4% PFA for 30 min. Afterwards, the membrane was blocked and incubated with a primary antibody (anti-α-syn 4D6, LB509, or anti-actin) and a secondary antibody conjugated with horseradish peroxidase. Protein bands on the membrane were detected by ECL-Plus detection system. Symbols (+) and (−) indicate presence and absence of membrane fixation with 0.4% PFA, respectively. Molecular size markers are shown in kilodaltons (kDa).