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Stereotypes are preconceived beliefs about individuals or groups, often based on incomplete or inaccurate information, leading to unfavorable attitudes and negative perceptions. These biases can result in discrimination, prejudice, and unfair treatment, causing harm to individuals and society. While gender-related biases and discrimination have decreased in recent years, gender stereotypes continue to hinder women’s career advancement. Negative gender stereotypes present significant challenges in human resource management, making it essential to address and reduce their impact, particularly the masculinization hypothesis affecting female employees. Although prior research has explored how stereotypes influence individuals, their effect on organizational performance remains understudied. Additionally, the role of femininity as a leadership trait in certain professions is poorly understood and requires further empirical investigation. This study focuses on how stereotype threat impacts individual outcomes, such as job performance. The research begins with reviewing relevant literature on stereotypes, stereotype threat, organizational environment, and performance, followed by developing a theoretical model and hypotheses. Using a sample of 160 female employees, the study employs questionnaires, reliability and validity tests, and SPSS software for correlation and regression analysis. It examines the direct relationship between negative stereotypes and workplace performance, the mediating role of stereotype threat, and the moderating effect of the organizational environment. Findings indicate that stereotype threat negatively affects professional women’s workplace performance. The study also highlights how negative gender stereotypes lead to biased judgments, hindering women’s career development and work quality. Furthermore, it investigates the effectiveness of social support systems in male-dominated environments, where women often face challenges related to gender identity. This research provides a comprehensive assessment of gender stereotypes, their impact, and theoretical implications, offering insights for mitigating their effects in organizational settings.