Departing Norfolk on 31 January 1942, in company with Emmons, Hambleton began a shakedown, unique in wartime, that took her through the Panama Canal to Callao, Peru; Valparaiso, Chile; Guayaquil, Ecuador; Cartagena, Colombia; and Balboa, Panama Canal Zone She was diverted for antisubmarine search north of Cuba in early March, and on 15 March 1942 rescued six men on a life raft who had survived the torpedoing of Ceibra.

After antisubmarine patrol along the East Coast and intensive training in Casco Bay, Maine, Hambleton sailed as escort to the cruiser Augusta and aircraft carrier Ranger on 14 April.

Reaching Africa's Gold Coast on 10 May, Ranger launched her cargo of Curtiss P-40s for the North African fighting and headed back to the West Indies.

With Royal Navy personnel on board as communications liaison, she conducted antisubmarine patrols and served as plane guard for the battleship Duke of York through August.

Hambleton joined the invasion fleet on 28 October, and as part of Admiral H. Kent Hewitt's Western Naval Task Force, she screened the escort carrier Sangamon during operations against airfields in French Morocco on D-Day, 8 November.

With all power gone, the destroyer took a 12 degree list to starboard as her damage control parties worked swiftly to jettison topside weights and shore up weakened bulkheads.

After a second shakedown in the Caribbean and training along the East Coast, Hambleton escorted a convoy to Oran in April 1944, and began to prepare for her role in the Normandy invasion.

She escorted a large convoy of LSTs to the landing areas on 7 June, D-Day plus 1, and remained off Omaha Beach for critical shore bombardment and screening duties.

Steaming via San Diego, Pearl Harbor, and Eniwetok, she arrived Ulithi 9 March 1945, to prepare for the invasion of Okinawa, the largest amphibious assault in the Pacific.

Departing Japanese waters 20 November, Hambleton steamed via Eniwetok, Pearl Harbor, and San Diego to Norfolk arriving late December.