The regiment was formed from independent militia companies operating in the Norfolk area, with men from the surrounding counties, as far west as Petersburg. Throughout the war it operated in brigades under William Mahone and David A. Weisiger, and the divisions of Benjamin Huger, Richard H. Anderson, and William Mahone. The regiment participated in the capture and later abandonment of Norfolk Naval Yard, and every major campaign of the Army of Northern Virginia. Several dozen men and officers of the regiment also served on the CSS "Virginia" in the Battle of Hampton Roads. The 41st Virginia was also involved in the friendly fire incident that severely wounded James Longstreet during the Battle of the Wilderness.