Background For better treatment outcome, exploring the knowledge of glaucoma and practices among glaucomatous patients is much more important. Objective The aimed of this study was to determine the knowledge and self-care practice and their associated factors towards glaucoma and its medications among glaucoma patients. Methods Cross section design study was conducted between May and July, 2017. The data was collected from glaucoma patients. Descriptive statistics was done to summarize socio demography data and associated factors with good knowledge and good practice were determined using binary logistic regression. Statistically significant variables were generated by considering p-value < 0.05. Results A total of 360 glaucoma patients were participated in this survey. The median age of the participant was 65 years. About 177 (49.25%) (95%CI: 43.9, 54.4) had good knowledge and a total of 197 or 54.7% (95%CI: 49.4, 59.7) had good practice about glaucoma and its medical treatments. The factors such as residence, marital status, educational status and stages of glaucoma of study participants were independently associated with having good knowledge. With respect to practice, educational status, duration of glaucoma, laterality and frequency of follow up were significantly associated with practice status of study participants. Conclusion In this study among glaucoma patients in a tertiary hospital, only half of patients have good knowledge about the disease and good practice pattern. Those with urban residence, higher education status and advanced stage of glaucoma were more likely to have good knowledge.