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The murder of the wealthy Sir Karell Borotyn in the study of his Czechoslovakian castle sparks renewed interest in a local legend that claims that the place is haunted by corpses that rise up at night to suck the blood of mortals.  Borotyn's body, found slumped over his desk, drained of its blood and marked with a distinct bite mark on the neck, is examined by Dr. Doskil, who quickly asserts that Sir Karell was killed by a vampire's bite.

After dismissing Doskil's conclusion as mere superstition, Inspector Neumann begins his investigation of the crime by questioning suspects Fedor Vincente, who stands to gain a large sum of money from Sir Karell's death as the fiancée of his only child Irena, and Baron Otto, Irena's new guardian and soon-to-be executor of her estate.  The investigation continues in a doctor's office, where the coroner questions a local innkeeper, who believes in the legend and says that the last time he saw the infamous vampires, Count Mora and his daughter Luna, they appeared as bats in the night.

When the learned Professor Zelin, an expert in the occult, learns of the circumstances surrounding Sir Karell's death, he, too, expresses his belief that Karell was killed by a vampire.  Next to encounter the mysterious forces of the castle legend is Fedor, who enters Irena's house in a state of obvious distress, unable to remember anything other than running by the old castle to catch a train then falling down.  Dr. Doskil attends to the fatigued Fedor, on whom he finds a bite mark that is identical to the one found on Sir Karell.

Later that night, a horse-led wagon rushes through the night carrying Maria, a new servant at the Borotyn estate, and as she and her driver pass by the old castle, they are halted by the frightening presence of Luna walking in the mist.  Maria returns to Irena's in a hysterical state, but Baron Otto insists that she merely had an encounter with real estate agents showing the property, and not the vampiress she claims to have seen.  When Irena is found inexplicably sitting in the garden in the middle of the night, it is suspected that she, too, has been attacked by Luna, and Professor Zelin orders that bat-thorn, a weed that wards off supernatural spirits, be spread throughout the household and that all the windows and doors remain shut.

Though it has been a year since Karell's death, his name is found on a recently signed lease for the castle, prompting an inspection of Karell's coffin, which is discovered to be empty.  While the help spreads more bat-thorn around the house, a bat flies in and is followed by the apparition of Count Mora.  After learning that the only way to rid the house of vampires is to find their graves during the day, slice their heads off and place bat-thorn in their necks, Inspector Neumann, Baron Otto and Professor Zelin form an expedition into the depths of the castle foundation to perform the deed.

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While sneaking up to one of the castle windows, Inspector Neumann and Baron Otto witness the strange sight of Sir Karell in the salon with Count Mora and Luna.  When Irena arrives at the castle and complains that she can no longer go through with Professor Zelin's plan to force Baron Otto's confession of her father's murder by frightening him with the sight of his ghost, it becomes apparent that the supposed supernatural occurrences of the past year were all part of a hoax designed by Zelin to crack Sir Karell's murder case.

Having failed in his effort to scare the baron with Karell's apparition, he then hypnotizes the man and, with the help of an actor posing as Sir Karell, initiates a re-enactment of the events leading up to the murder.  The conversation that the two men had prior to the murder reveals that the baron forbade Karell's consent to the marriage of his daughter to Fedor.  Having secured the motive of the crime, Zelin and Neumann observe the baron as he prepares to murder the actor posing as Karell, but stop him once they discover his blood-letting technique.  Irena apologizes to Fedor for not letting him in on the scheme, and Count Mora and Luna remove their costumes and plan their next stint as part of their haunted theater act.

Notes
While MPH claims that this film was based on a novel entitled "The Vampires of Prague" by Guy Endore and Bernard Schubert, modern sources indicate that Mark of the Vampire was a remake of the the 1927 MGM silent film London After Midnight, which was based on Tod Browning's story "The Hypnotist."  It has not been determined which is correct.  Working titles for the film were Vampires of Prague and Vampires of the Night.

HR production charts list actors Jane Mercer, Henry Stephenson and Doris Lloyd in the cast, but their appearance in the released film has not been confirmed.  A contemporary NYT news item notes that MGM imported a number of large South American bats for the picture, and was ordered by the government to either deport or destroy the creatures after the film was completed.  According to modern sources, Rita Hayworth was tested for the part played by Carol Borland.  Mark of the Vampire marked the screen debut of Borland, who, according to a Bela Lugosi biography, appeared in a stage production of Dracula with Lugosi, but concealed her prior professional involvement with the actor while testing for the role.

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Lugosi's biography also notes that the actor designed his own costume, and that neither he nor any of the other actors knew how the film was going to conclude until the final days of production, when Browning made the final pages of the script available to them.  Because the actors had been playing the story as strict horror, they reportedly balked at Browning's "gimmick" ending.  An alternate ending with a second twist, in which Lionel Barrymore's character receives a telegram from the vaudeville actors apologizing for not being able to make their train for the castle assignment, was proposed, but Browning rejected it.  The original story had Count Mora committing suicide after killing his daughter, with whom he had an incestuous relationship, but all traces of the incest and suicide plots, with the exception of Count Mora's bullet wound scar resulting from the suicide, were removed from the film.

Modern sources list Jack Dawn and William Tuttle as the makeup artists.

Earlier reviews list running times in excess of eighty minutes, a possible indication that the film was cut after previews.

Following the release of Mark of the Vampire, the NYT printed a letter it received from a physician who wrote to the Screen Editor of the paper to complain that "a dozen of the worst obscene pictures cannot equal the damage that is done by such films as The Mark of the Vampire."  Robinson objected to the film from a medical standpoint, stating, "I refer to the terrible effect that it has on the mental and nervous systems on not only unstable but even normal men, women and children...several people have come to my notice who, after seeing that horrible picture, suffered nervous shock, were attacked with insomnia, and those who did fall asleep were tortured by most horrible nightmares."  The letter concluded: "In my opinion, it is a crime to produce and to present such films.  We must guard not only our people's morals—we must be as careful of their physical and mental health."  According to the MPAA/PCA Collection at the AMPAS Library, Mark of the Vampire was rejected by censors in Poland.  In Hungary, censors deleted all screams throughout the picture, as well as shots of bats, spiders and "the more gruesome shots of the vampire."