Background Voluntary blood donation is critical for maintaining an adequate and safe blood supply. Despite awareness campaigns, there remains a disparity between the demand and actual voluntary blood donations in India. Understanding the knowledge, attitudes, practices (KAP), and influencing factors among voluntary donors can aid in developing targeted interventions, particularly in regions like Western Maharashtra where such data is limited. Aims and Objectives The primary aim of this study was to assess the knowledge, attitude, and practices regarding blood donation among voluntary. The study also aimed to explore barriers and motivators influencing donor behaviour. Methods A cross-sectional descriptive study was conducted among 400 voluntary blood donors attending selected blood donation camps and hospital blood banks in Western Maharashtra. Data were collected using a pretested, semi-structured questionnaire covering KAP domains and sociodemographic details. Descriptive statistics were used for data summarization. Chi-square test and logistic regression were employed to determine associations between KAP levels and independent variables. Results Among participants, 56% had average knowledge, 10% demonstrated good knowledge, and 34% had poor knowledge. Positive attitudes were seen in 68% of donors, while only 44% had favourable practices (e.g., regular donation). Education level and gender were significantly associated with knowledge and practice (p < 0.05). Major motivators included altruism and peer influence, while fear of weakness and lack of awareness were key deterrents. Conclusion The study revealed gaps in knowledge and practices despite favourable attitudes among donors. Educational status and gender significantly influenced donor behaviour. Tailored awareness campaigns, especially targeting less-educated groups and males, may enhance voluntary donation rates.