Comparison of plasma prefractionation methods using SDS-PAGE and Western blot analysis. (a) 12.5% SDS-PAGE (10 μg/lane) showed consistency and reproducibility of protein band patterns with thermal treatment as well as with untreated plasma, IMDP, and CPLL. Protein bands were visualized by the blue silver CBB-G250 staining . The interindividual variable proteins, so-called the Band-A, were labelled in the red square. (b) Western blot analysis (10 μg/lane) demonstrated that three methods could deplete HSA and IgG, the most abundant plasma protein, as compared to the untreated plasma and thus met a required characteristic for plasma prefractionation method. Full-length immunoblots were provided in Supplementary Figure 1. Plasma samples from three subjects (S1-S3) were used for both experiments, while three technical replicates per specimen were also performed for SDS-PAGE.