Santana had his first taste of WWF success in 1979 when he teamed with Ivan Putski to defeat Johnny Valiant and Jerry Valiant for the WWF Tag Team Championship at Madison Square Garden in October 1979. The duo held the titles for close to six months before losing to the Wild Samoans in April 1980.
In 1983, he engaged in a lengthy feud with Intercontinental Champion Magnificent Don Muraco. Santana finally won the title on February 11, 1984, becoming the first Mexican-American wrestler to win the WWF Intercontinental Championship. He quickly entered into a feud over the Intercontinental Championship with Greg Valentine. Valentine captured it from Santana in September 1984 in London, Ontario. Soon after, the plot had Valentine injuring Santana's knee and putting Santana out of action for several months.
Santana returned in December 1984 and set his sights on getting the Intercontinental title back from Valentine. During this time he started using The Hammer's finishing hold, the Figure four leglock and also wrestled in tag-team competition with Blackjack Mulligan. Santana wrestled at the first ever WrestleMania at Madison Square Garden in March 1985, and in the opening match defeated a masked wrestler known as The Executioner ("Playboy" Buddy Rose), making him submit to the figure four in 4:05. Santana made an appearance in the ring later in the card during the IC Championship match between Valentine and the Junkyard Dog, Santana's friend. Wearing street clothes, Santana rushed to the ring to inform referee Dick Kroll that Valentine had used his feet on the ropes to help pin JYD. Despite having already called for the bell, Kroll restarted the match and Valentine was counted out as he didn't get back into the ring to continue.
Santana and Valentine went on to wrestle a memorable series of singles and tag team matches with neither gaining the upper hand. They wrestled in a variety of different types of matches such as regular title matches, No Disqualification matches, and Lumberjack matches.
Santana recaptured the Intercontinental title from Valentine in a brutal steel cage match in Baltimore on July 6, 1985. Valentine, incensed over losing the belt, destroyed it by repeatedly bashing it against the steel cage. This forced the WWF to get a new Intercontinental title belt (in reality, the WWF had made a new IC Championship belt to go along with the new image they were trying to promote and 'smashing' the old IC belt was seen as a way of moving forward with Santana being the first to wear the new belt). Tito would hold onto the title until February 8, 1986 when he lost it to Randy Savage at the Boston Garden after Savage knocked him out with a foreign object that went unnoticed by referee Danny Davis.
Santana formed the popular tag team Strike Force with Canadian Rick Martel in the late 1980s. They defeated the Hart Foundation for the WWF World Tag Team titles and held them for five months before losing to Demolition at Wrestlemania IV. Due to a neck injury inflicted on Martel (kayfabe) shortly after the loss, the team was inactive for several months. Immediately after the injury, Santana introduced a new tag team to the WWF, the Powers of Pain, whom he briefly managed. The Powers were introduced as mercenaries to help Martel and Santana gain revenge on Demolition for both the title loss and the injury to Martel. The Powers would later find more permanent management with The Baron before finally turning heel at the 1988 Survivor Series by stealing away Demolition's manager Mr. Fuji, leaving the champions as babyfaces.
Martel returned at the Royal Rumble in 1989 and reunited with Santana. However, in their WrestleMania match against the Brain Busters (Tully Blanchard and Arn Anderson), Martel turned on Santana during the match after accidentally being hit by Santana's Flying forearm smash. Martel refused to tag in and walked back to the dressing room leaving Santana to face both opponents alone (the Busters then easily defeated Tito with a Spike piledriver). In an interview with "Mean" Gene Okerlund immediately following the match, Martel called Santana a loser and said he was sick and tired of carrying him. His feud with the newly heel Martel would last throughout 1989, with both men on opposing teams at both SummerSlam and Survivor Series and Santana defeating Martel in the finals of the 1989 King of the Ring tournament. Tito even allied with his former archenemies Demolition against Martel, defeating him and The Fabulous Rougeaus in a six man tag match on June 22, 1989 in Hartford, Connecticut.
After the Ultimate Warrior won the WWF Championship from Hulk Hogan at WrestleMania VI and vacated the Intercontinental Title, Santana took part in an eight-man tournament to name a new Intercontinental Champion. Santana made it to the finals, where he lost to Mr. Perfect. Following that loss, Santana occasionally teamed with fellow fan favorite, Koko B. Ware.
At the 1990 Survivor Series, he teamed with Nikolai Volkoff and The Bushwhackers. He was the winner and sole survivor in the elimination-style match against Sgt. Slaughter, Boris Zhukov, and The Orient Express. As a result, Santana advanced to the final elimination match, teaming with Hulk Hogan and the Ultimate Warrior against Martel, Ted DiBiase, the Warlord, and Power and Glory. Santana would eliminate the Warlord before being pinned by DiBiase. Santana would then wrestle at WrestleMania VII, losing to The Mountie in a little over a minute.
Santana then adopted a Spanish bullfighter gimmick and the nickname "El Matador" in 1991. Under this gimmick, he faced Shawn Michaels in the opening bout of WrestleMania VIII at the Hoosier Dome in Indianapolis, Indiana. Santana wrestled under the "El Matador" gimmick through 1993, mostly as a jobber to the stars. This included a loss to Papa Shango at SummerSlam (1992). In his final in-ring WrestleMania appearance, he defeated Shango at WrestleMania IX in the untelevised opening match.
In his final appearance on WWF programming, Santana defeated friend and frequent tag team partner Virgil on a 1993 episode of Wrestling Challenge. As a sign of mutual respect between the two, both men embraced after the match.
Santana returned to the WWF as a commentator in the Spanish Broadcast table in 1997. He called on Monday Night Raw, as well as PPV events. He was last doing Spanish commentary at WrestleMania XIV.
On March 13, 2004 he was inducted to the WWE Hall of Fame, with an induction speech by his WrestleMania VIII opponent Shawn Michaels.
On the November 15, 2010 "Old School" episode of Raw, he ring-announced Alberto Del Rio.