The focus of the show (which was initially entitled The Witch of Westport according to the A&E Network Biography of Elizabeth Montgomery), is on the mixed marriage of a nose-twitching witch, Samantha Stephens (Montgomery), and her mortal husband, Darrin (originally played by Dick York and later by Dick Sargent). The series, a romantic comedy, shows how true love can endure the most vexing of situations, even between a witch and a human. Samantha's mother, Endora (Agnes Moorehead), disapproves of Darrin, as indeed do most of Samantha's family. Many episodes revolve around Endora, or another of Darrin's spell-binding in-laws, using magic to undermine the union. Although Endora casts countless spells on Darrin, she never outright banishes him from Samantha's life, or casts any spells of permanence. Endora's ploy seems to be to provoke Darrin into leaving Samantha, but the mortal's love for his wife overcomes every obstacle. Endora refuses to even remember Darrin's name, invariably calling him "Durwood", "Darwin," and even "Dum-Dum," much to his annoyance.

Most episodes began with an enraged Darrin becoming the victim of spell. By the epilogue, however, Darrin and Samantha would embrace and confound the devious elements that failed to separate them. Some storylines took a backdoor approach to such topics as racism, as seen in the first season episode, "The Witches Are Out". Samantha objects to Darrin's demeaning portrayal of witches as ugly and deformed in a Halloween candy ad. Such stereotypical imagery, she believes, ignites biases which have often caused Endora and her to flee the country until the season ends. One 1970 in television episode, "Sisters of the Heart", was written and submitted by a tenth grade class. It involved Tabitha altering her and her black friend's skin tones with coordinating polka-dots, so that people would treat them alike (a similar white-with-black vs. black-with-white concept was seen earlier on a Star Trek episode that featured Frank Gorshin).

Bewitched was unique for the mid-1960s in that it portrayed an estranged married couple - Samantha's parents, Endora (Agnes Moorehead) and Maurice (Maurice Evans). Maurice was portrayed as an urbane thespian not unlike Elizabeth Montgomery's father, Robert Montgomery. Maurice occasionally pops in with a young, attractive female escort. The couple's separation is inferred and subtextual. In the episode, "Samantha's Good News", Endora threatend to file for an "ectoplasmic interlocutory" (code for divorce), only to wrangle Maurice's affection.

Darrin works for advertising agency McMann and Tate, and his profit-obsessed boss Larry Tate (David White) is an almost constant presence on the show (though Tate's partner McMann appears only twice during the series). Tate's opinions would turn on a dime to appease a client and land a deal. Despite witnessing plenty of oddities, Larry and his wife Louise (Irene Vernon, and later, Kasey Rogers) never discover that Samantha is a witch. Agnes Moorehead as Endora. Agnes Moorehead as Endora.

The sole member of Samantha's family for whom Darrin shows any affection is the lovably bumbling Aunt Clara (played by Marion Lorne). Lorne won a posthumous Emmy in 1968 for the role of the aged witch, whose powers have begun to wane, and whose spells often end in disaster. She appeared in 27 episodes; when Lorne died during the fourth season, the absence of Aunt Clara was left unexplained. Because Lorne was so popular and loved by the cast, another actress taking the role was not considered. A similar character, the anxiety-ridden witch-nanny Esmeralda, played by Alice Ghostley, would later make appearances starting during the 1969-1970 season. (Both Ghostley and comic actor Paul Lynde had guest roles as mortal characters before being cast as magical semi-regulars.)

Samantha's far-out and egocentric lookalike cousin Serena is another important character, first appearing in season two. The character, also played by Elizabeth Montgomery, would be credited as "Pandora Spocks" from 1968-72. Ever mischievous, Serena often chases after Darrin and Larry Tate (calling the white-haired Tate "Cotton-Top"), just for fun. Serena is the antithesis of Samantha, sporting a heart-shaped birth mark on her cheek, raven black hair, and mod mini-skirts. More progressive than the typical witch or warlock who generally loathes mortals, Samantha's counter-culture cousin occasionally dates some (including a character played by Jack Cassidy).

During the show's run, both Aunt Clara (to Louise Tate) and Serena (to Phyllis Stephens) state they are from Maurice's side of the family, though Serena sometimes plots with "Auntie" Endora. Despite her wild behavior, Serena ultimately supports Samantha and Darrin, even though she finds them both a bit square.

Endora's prank-loving brother Arthur (Paul Lynde) makes several memorable appearances. In one episode, Arthur befriends Darrin and teaches him a phony ritual that will set Endora straight. Darrin performs the silly ritual to the outright bewilderment of his wife and mother-in-law. After Arthur erupts in rip-roaring laughter, it becomes clear that Darrin was set up. The trio get even with Arthur when Darrin's own practical joke seemingly obliterates Endora. Despite the hoax, Arthur genuinely likes Darrin. In another episode, Serena and Uncle Arthur go toe-to-toe with the Witches' Council to support the Stephens union, only to have their powers suspended.

Veteran actress Mabel Albertson (sister of Jack Albertson) plays Darrin's straight-laced mother, Phyllis. The character often complains of "a sick headache" when she witnesses a magic spell in motion. Adding to the fun are the Stephenses' witch daughter Tabitha (Erin Murphy), and her baby brother Adam, as well as various witches, warlocks, and mere mortals, along with an occasional elf, leprechaun, nymph, and other supernatural being. The program made full use of the period's modest but effective special effects wizardry.

Across the street lives a retired couple, Abner Kravitz (stage and movie actor George Tobias) and his nosy wife, Gladys. Gladys was played first by Alice Pearce, who won a posthumous 1966 Emmy for the role; following Pearce's death in 1966 from ovarian cancer, the character was played by Sandra Gould. On the studio backlot, the Kravitzes' house was actually down from the Stephens' house exterior. Both homes' exterior doors opened to an unfinished ten-by-fifteen foot entry, as the interiors were shot elsewhere. The exterior of the Kravitzes' house later became the home of The Partridge Family.

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