When it started, it featured three or four stories of horror and suspense in each issue. With issue #143 (June 1964), the Martian Manhunter series moved over from Detective Comics. He would continue here until #158 (April, 1966).
With issue #156 (January, 1966), Dial H for Hero begins and is the cover feature until issue #173 (March-April 1968). At this point, House of Mystery returns to being a true horror-mystery anthology title.
With #175 (May-June 1968), almost all the tales were introduced by a character named Cain, 'able care taker' of the House of Mystery, who was also one of the hosts of Plop! and who later became a recurring character in The Sandman. The horror format began with the previous issue, including a character resembling the Phantom Stranger. Four issues from this era, #s 260, 267, 268, and 274, feature stories by T. Casey Brennan.
Its most famous recurring feature was I...Vampire, which dealt with a heroic vampire, Andrew Bennett, who sought to defeat his nemesis and former lover Mary, the Queen of Blood. This series began in #290 (March, 1981) and would last until #319 (August, 1983), two issues before the title ended with #321 (October, 1983).
The series won a good deal of recognition in the comics industry, including the Shazam Award for Best Individual Short Story (Dramatic) in 1972 for "The Demon Within" in #201 by John Albano and Jim Aparo, and the Shazam Award for Best Humor Story in 1972 for "The Poster Plague" by Steve Skeates and Sergio Aragones.
In its last years, the series was edited by Karen Berger, who ultimately founded the Vertigo imprint. this series is generally regarded as one of Vertigo's roots.