2.5. Western Blot Analysis

Small nuclear ribonucleoprotein polypeptide A (SNRPA) protein was expressed and purified from yeast as GST fusions by CDI Labs. The easily purified protein was separated by electrophoresis on sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (EZBiolab) and then electrotransferred onto poly(vinylidene fluoride) membranes. After blocking with 5% BSA in PBST, the membranes were incubated with serum (1:200) for 2 h. Membranes were washed three times with TBST, followed by incubation with HRP-conjugated goat anti-human IgG (EASYBIO; 1:5000) for 1 h. After washing with TBST and deionized water, the immunoreactive bands were detected by the sensitive chemiluminescence substrate (Thermo Fisher Scientific) and visualized using Clinx Chemical Capture (Science Instruments).