This survey research article examines the evolving landscape of women’s political participation and representation in South Sudan during the critical implementation phase of the Revitalised Peace Agreement (2021–2026). It addresses a significant gap in contemporary African political studies by investigating the lived experiences and perceived barriers faced by women navigating formal political structures in a post-conflict state. Employing a sequential mixed-methods design, the study analyses quantitative survey data from 450 women across five states, triangulated with qualitative insights from focus group discussions with 60 female political aspirants and elected officials. Key findings reveal that, despite constitutional provisions for a 35% representation quota, substantive participation remains constrained by entrenched socio-cultural norms, limited access to financial resources for campaigning, and systemic party-level gatekeeping. However, the data also indicate a resilient cadre of women strategically leveraging informal networks and civil society platforms to influence policy agendas. The study concludes that achieving meaningful gender parity requires moving beyond nominal quotas to dismantle the foundational socio-economic and institutional barriers that perpetuate exclusion. These findings contribute crucial empirical evidence to African feminist scholarship and offer direct implications for policymakers and civil society organisations designing interventions to strengthen women’s political agency in South Sudan and comparable post-conflict settings.