The second season of American Ninja Warrior began on December 8, 2010, in Venice Beach, California where 300 competitors took on the course. The 15 semi-finalists moved on to the "Ninja Warrior Boot Camp" in the remote California mountains, where they competed in a series of team challenges. Then, the final moved on to compete in the season finale of American Ninja Warrior 2 as a part of Sasuke 26 at Mount Midoriyama in Japan. The series concluded on January 2, 2011, same as Sasuke 26 was aired in Japan.

This season was hosted by Matt Iseman and Jimmy Smith, with G4's Alison Haislip as a sideline reporter.