Screenshot/YouTube
Trump famously went to war with the Khan family earlier this year over their critique of his comments about veterans and proposal to bar Muslims from entering the US.
The Republican presidential nominee criticized Khizr Khan, Humayun's father, for appearing in multiple television interviews following his speech at the Democratic National Convention.
The new Clinton campaign ad opened with footage of Khan arranging his son's uniform and talking about his sacrifice in Iraq.
"In 2004, my son was stationed in Iraq. He saw a suicide bomber approaching his camp," Khan said. "My son moved forward to stop the bomber when the bomb exploded."
He continued: "He saved everyone in his unit. Only one American soldier died. My son was Captain Humayun Khan. He was 27 years old, and he was a Muslim American."
The ad ended with Khan asking, "Mr. Trump, would my son have a place in your America?"
The ad is airing in Florida, Iowa, Nevada, New Hampshire, North Carolina, Ohio and Pennsylvania.