Ortiz garnered widespread attention for her role as Hilda Suarez in the ABC comedy-drama series Ugly Betty from 2006 to 2010. She also appeared such films like Labor Pains (2009) and Big Mommas: Like Father, Like Son (2011), and starred in Little Girl Lost: The Delimar Vera Story (2008). In 2013, Ortiz began starring as Marisol Suarez in the Lifetime television comedy-drama series Devious Maids, for which she received the Imagen Award for Best Actress - Television.
Ortiz was born in Manhattan, and is the daughter of Angel L. Ortiz, a former Philadelphia City Council member of Puerto Rican descent, and an Irish American mother. As a child, Ortiz originally dreamed of becoming a ballerina, and she studied ballet for eight years, until the pain of dancing en pointe forced her to pursue a different artistic discipline. Ortiz graduated from The University of the Arts in Philadelphia.
Switching to singing, Ortiz attended the Fiorello H. LaGuardia High School of Music & Art and Performing Arts in New York City and later the University of the Arts in Philadelphia before making her professional stage debut in a regional theater production of Dangerous Liaisons. Her additional stage credits include portraying Chrissy in a European touring production of Hair, regional theater productions such as Dog Lady and the Cuban Swimmer and in the South Coast Repertory Theater's production of References to Salvador Dalí Make Me Hot. After joining LAByrinth Theatre Company, she appeared off-Broadway in In Arabia, We'd All Be Kings, which also co-starred Puerto Rican actress Liza Colón-Zayas and was directed by Academy Award-winning actor Philip Seymour Hoffman and named one of the 10-best plays of 1999 by the magazine Time Out New York.
Ortiz appeared in several television shows including NYPD Blue, Everybody Loves Raymond, Commander In Chief, ER, and The New Adventures of Old Christine. She had her first series regular role in short-lived NBC comedy series Kristin (2001), opposite Kristin Chenoweth. She later starred in another short-lived sitcom, called A.U.S.A. in 2003, and later had recurring roles in the FX drama Over There, and ABC series Boston Legal.
In 2006, Ortiz was cast as Hilda Suarez, the lead character's sister, in the ABC comedy-drama series Ugly Betty. Originally, Ortiz had auditioned for the Betty Suarez role, but it went to America Ferrera instead. Ortiz said in 2008, "I was super plain. I just put on some glasses and kept my hair kind of flat and layered on the clothing", describing what she wore when auditioning for the role of Betty. "I just wanted them to remember me, so that I could be involved in the future". Ortiz was one of two Philadelphians who were regulars on Ugly Betty, the other being Mark Indelicato, who played her son Justin. Another connection to the city is co-star Becki Newton (who played Amanda Tanen), who graduated from the University of Pennsylvania in Philadelphia.
In addition to her role on Ugly Betty, Ortiz was active in films, which include starring roles Tortilla Heaven, and Labor Pains co-starring her former Ugly Betty co-star, Lindsay Lohan. She also had a voice role in Batman: Gotham Knight, and starred in the made-for-television movie Little Girl Lost: The Delimar Vera Story in 2008. Ortiz also had a guest-starring role on Army Wives during its second season. She played a waitress named Sandi. She also appeared in the 2010 music video for Enrique Iglesias and Juan Luis Guerra's "Cuando me enamoro".
After Ugly Betty was ended in 2010, Ortiz was cast as lead character in ABC drama pilot True Blue about San Francisco homicide detectives reunited to solve the murder of one of their own. The pilot was not picked up to series. In next year she appeared in the comedy film Big Mommas: Like Father, Like Son, and starred in another pilot, called Outnumbered for Fox. This pilot also was not picked up to series. She later had a recurring role of HBO comedy series Hung. Ortiz also starred in the independent films Sleeping with the Fishes, and Such Good People.
In 2012, Ortiz was cast as lead character of ABC comedy-drama pilot Devious Maids, created by Marc Cherry. On May 14, 2012, the pilot was not picked to series by ABC for the 2012–13 United States network schedule. However, on June 22, 2012, Lifetime picked up the pilot with a thirteen-episode order. The series premiered June 23, 2013, on Lifetime with positive reviews from critics. In show, she plays the lead role of Marisol Duarte (the alias of Professor Marisol Suarez). Interestingly, this will be the second series in which her character shares the last name of her previous character from another series. Also in 2013, Ortiz was special guest star in ABC primetime soap opera, Revenge, as Bizzy Preston. In 2014, Ortiz guest-starred in the two another ABC shows: How to Get Away with Murder produced by Shonda Rhimes, and Black-ish.
On October, 2014, Ortiz was cast alongside Katherine LaNasa, Jeremy Sisto and Tyler Blackburn in the gay-drama, Love is All You Need?, based on the 2011 short film with the same name.
In an interview she was seen saying that she is incredibly proud of her Devious Maids.
Ortiz currently lives in Los Angeles. In her spare time she enjoys hiking, horseback riding, softball, and water sports, and writes poetry. She married musician Noah Lebenzon on June 9, 2007 in Rincón, Puerto Rico. The couple was expecting their first child on June 25, 2009. On Saturday, June 27, 2009, Ana and Noah welcomed a daughter, named Paloma Louise Lebenzon. Paloma was born at 4:25 p.m. and weighed in at 7 pounds 10 ounces. The second child, a son named Rafael, was born on September 24, 2011.
Ortiz is active in raising awareness against domestic abuse, citing her own experiences when she was in her 20s and involved in a romantic relationship that turned physically abusive. In an article for USA Today, Ortiz notes that the final scene from the Ugly Betty episode "How Betty Got Her Grieve Back", in which Hilda sat on her bed staring at the bathroom after realizing that Santos had died, was drawn from that experience: "I used some of that subconsciously ... It's something that is always with me".