The study aimed to optimise post-activation potentiation (PAP) strategies for Rider 1 in elite team sprints to improve performance over 250 m (opening lap), with a focus on female cyclists. Eight national-level track cyclists participated in this study, undergoing four sets of activation strategies: control (CON), dynamic high inertia (DYN, 4 × 4 pedal strokes), isometric contraction (ISO, 4 × 4 s, 4 angles), and back squat activation (BSQ, 4 × 4 rep, 80%1RM). The tests were divided into pre-activation and post-activation phases, including measurements at 4 min, 8 min, and 12 min after activation. The tests included a 250 m time trial (TT) and segment timing, with measurements of peak torque, peak power, average power, and cadence. The mean cadence, torque, and power for the first 62.5 m of pedal revolutions were collected. Paired-sample t-tests were used to assess activation differences. Multiple group comparisons were conducted using analysis of variance (ANOVA). The Bonferroni correction was used to control Type I errors. For significant activation strategies, linear or non-linear regression was applied to extrapolate the torque-cadence and power-cadence profiles, and the parameter differences were examined to investigate profile changes. Cohen’s d and Cohen’s f were used as effect sizes. After DYN activation, the 250 m TT significantly improved (p = 0.018), primarily through a reduction in the 62.5 m time (p = 0.006) and an increase in peak torque (p = 0.018). After 12 min of ISO activation, the 250 m TT showed a large effect but did not reach the significance level under Bonferroni correction (p = 0.135, d = 0.860), with a notable reduction in the 62.5 m time (p = 0.003). PAP can be strategically employed to enhance the performance of elite female Rider 1 in team sprints.