At the age 32, Yoon Sang-hyun made his acting debut in the 2005 SBS series Marrying a Millionaire. The following year, he bagged the New Star Award at the SBS Drama Awards for his performance in the drama Common Single. His breakout role came in 2009 when he played an immature chaebol in the hit MBC romantic comedy Queen of Housewives. His work in the series won him a Top Excellence Award at the 2005 MBC Drama Awards. He also scored a Best Actor nomination at the 46th Baeksang Arts Awards for the series. Also in 2009, he headlined the KBS2 drama My Fair Lady, in which he took home the Popularity Award as well as the Best Couple trophy with Yoon Eun-hye at the KBS Drama Awards. In 2010, he joined the main cast of the top-rating SBS series Secret Garden, which was hailed Best Drama at the 47th Baeksang Arts Awards. In 2018, he was paired with Han Hye-jin in the MBC romantic melodrama Let’s Hold Hands and Watch the Sunset, in which he was nominated for a Top Excellence Award at the MBC Drama Awards.

As a singer, Yoon Sang-hyun contributed his vocals to the official soundtracks of some of his dramas. In 2011, he released his first full-length album in Japan titled Precious Days.

Yoon Sang-hyun tied the knot with singer MayBee on February 8, 2015. They have two daughters and one son together.