The ad from Our Principles PAC, a top anti-Trump super PAC, will air in the final five-day stretch ahead of Tuesday's crucial Ohio primary. It comes in addition to the more than $3 million the group is spending in Florida and Illinois, said Tim Miller, a spokesman for the PAC and who used to work for the Jeb Bush campaign.
The 30-second TV spot, shared first with CNN on Thursday, intersperses clips of Trump rallies lamenting the lack of "made in the U.S.A." labels with a clip of the billionaire businessman getting taunted by late-night talk show host David Letterman over where Trump's line of clothing is made.
"Remember we used to have made in the USA, right? When was the last time you've seen it," Trump says in a first clip pulled from a recent rally.
The ad then shows then-CBS' "Late Show" host Letterman asking Trump where his products are made.
"The ties are made in, where, China?" Letterman asks in an October 2012 interview as he holds several ties from the Donald J. Trump Collection.
After Letterman reveals that a collared shirt from Trump's clothing line is made in Bangladesh, Trump says, "Well, that's good, we employ many people in Bangladesh."
Many of the clothes from Trump's clothing line are made in foreign countries including China and Mexico, the two countries Trump constantly says are "killing us on trade."
On the campaign trail, Trump has repeatedly called on American companies, including Nabisco, Ford and Apple, to stop outsourcing manufacturing jobs and has vowed as president to impose high tariffs on American companies that try and import products manufactured abroad.
"It's very hard to have apparel made in this country," he has said, also hitting China for manipulating its currency.
The TV ad will also be supported by a digital advertising and direct mail campaign in Ohio that will also hit Trump's record on outsourcing.
Miller, who served as communications director to Bush, said donors to the super PAC did not request ad placements in Ohio, but new cash injections provided sufficient funds for the group to make a dent in the key primary state.
"We wanted to make sure that every place we went we could have an impact. These are not show buys, these are targeted," Miller said.
He predicted that the Ohio race -- where different polls show Trump and John Kasich are vying for the lead -- will be close and that the group anticipates boosting its paid media efforts in Ohio ahead of Tuesday's winner-take-all primary.