Call of Juarez: Bound in Blood was aimed to make the player's experience as the "best West ever created".^[8] The game derives from spaghetti-western genre: gunslingers, outlaws, duels, prison breaks, bank robberies, stagecoach chases and conflicts with Native Americans, similar to another western game, Gun. It features two characters, brothers Ray and Thomas. The game plot is set in the American Civil War period and presents the adventures of two brothers across different parts of North America.Before most missions, the player can choose between one of the two brothers, but sometimes only one is available.

Though the game's Story Mode is fairly short, each brother has a different effect on the missions: often the player will run into split paths for either brother to take, or certain key points where one brother needs to do particular things while the other does something else. For example, Thomas can lasso on top buildings for a vantage point to cover Ray while he attacks/dynamites certain areas. One brother's story mode will be quite different from the other's. At first the game just moves from mission to mission but once the game opens, between certain missions there will be an open Free World where the player can take side-missions as a bounty hunter or hired gun, respectively for either attack or defence, and other kinds of side-missions. In each mission and each Free World there will be a trader who will sell the player certain weapons available at different qualities.