Lumpy skin disease virus (LSDV), a member of the capripoxvirus genus, induces lumpy skin disease (LSD) in cattle and buffalo populations. The initial documentation of LSD dates back to 1929 in Zambia, with subsequent rapid dissemination within the cattle community in Africa and Asia. Described as the “smallpox” equivalent in cattle, LSD manifests through clinical features such as fever, extensive cutaneous nodules, body wasting, lymph node enlargement, and skin edema. Isolation of LSDV constitutes an important step in evaluating virulence and implementing disease control measures. Skin samples obtained from afflicted cattle serve as the primary source for LSDV isolation due to its tropism for epidermal cells. Other tissues, including blood, lung, spleen, and saliva samples, are also employed for viral isolation. This chapter presents protocols for the convenient isolation of LSDV, relying on the plaque formation phenotype and utilizing MDBK cells. Emphasis is placed on stringent quality control methodologies to identify isolated viruses through multiple techniques, facilitating further investigations such as the in vitro growth of isolated LSDV.