Stanton ended a 10-game homerless drought and became the 147th player to reach the mark, 11 of them active. He did it in 1,119 games, making him the fifth fastest to hit the milestone behind Ralph Kiner, Ryan Howard, Juan Gonzalez and Alex Rodriguez.
At 28, Stanton was the ninth-youngest player to hit 300. He hit his 33rd homer for New York, who lost the game 8-7. He led the majors with 59 last year and was the NL MVP before Miami traded him to the Yankees.
Stanton went the opposite way for a drive into the right-field seats on a 3-1 pitch from left-hander Francisco Liriano.
Stanton watched the path of the ball for a couple of seconds before flipping his bat aside with a small smile on his face. After rounding the bases and receiving a warm ovation from his teammates, he emerged from the dugout and doffed his helmet to the fans offering a standing ovation.
The home run continued a gradual feeling-out process between the fans and Stanton, whose slumps have made him the occasional target for boos during his first season with the Yankees. He has had three dry spells of 10 or more games without a home run this year.
Information from The Associated Press contributed to this report.