Disuccinimidyl suberate (DSS) and disuccinimidyl sulfoxide (DSSO) exemplify these two strategies, respectively. Conventional wisdom suggests that DSS is optimal for simple samples, while DSSO excels in complex proteomes. However, this widespread assumption lacks rigorous, systematic validation. Here, we present a direct, side-by-side performance evaluation of DSS and DSSO across low-, medium-, and high-complexity protein samples.