Introduction: Insulin is essential for the normal carbohydrate, protein and fat metabolism in the body. There has been emphasis on early initiation of Insulin in routine diabetes care. Insulin therapy presents many challenges due to complexities associated with its intricate use. An inadequate knowledge of its use can result in preventable complications, adverse patient outcome, poor adherence to therapy and invariably poor glycaemic control. This study aimed at determining the level of awareness among the Insulin taking populations, both urban and rural, about Insulin and the methods of Insulin administration, storage and disposal of syringe. Method: This study was a cross-sectional study conducted on100 participants with Type 2 diabetes mellitus (T2DM) who took inpatient or outpatient care under Department of General Medicine, St Johns’ Medical College Hospital, Bangalore, India. A self-administered standardized questionnaire was used to collect the data. The tool assessed knowledge on administration, storage, usage and disposal of Insulin or insulin syringe. With proportions, mean & standard deviation, details of knowledge of insulin use was classified as Adequate and Inadequate knowledge. Result: The hundred participants consisted of 59 males and 41 females who were present in a tertiary care hospital in Bangalore, south India. The majority of population belonged to urban/ periurban background. Most participants 57(57%) were found to have inadequate knowledge with regard to administration, storage and usage of Insulin. Poor disposal technique was present among 82% of the study population. Conclusion: The knowledge regarding Insulin use - administration, storage, usage and disposal among the Insulin using participants was inadequate, indicating the need for better awareness programs by health care professionals before starting insulin among patients.