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The advert also features a statement from Secretary of State Hillary Clinton at a press conference rejecting the amateur film's message.
Unrest over the film, Innocence of Muslims, has claimed several lives.
Also on Thursday, a protest against the film outside the US embassy which had turned violent ended peacefully.
The adverts seek to emphasise the message reiterated by US officials throughout the crisis: that the "disgusting" film was not made by the US government, but that there is never any justification for violence.
A caption on the advert reads "Paid Content," the Associated Press reports, although the embassy itself refused to confirm whether or not the advert had been paid for.
The embassy described the advert as a "public service announcement" and repeated the statements from Mr Obama and Ms Clinton on its Twitter feed.
The low-budget film that sparked the controversy was made in the US and is said to insult the Prophet Muhammad.
Its exact origins are unclear and the alleged producer for the trailer of the film, Nakoula Basseley Nakoula, is in hiding.
The Pakistani authorities had earlier called on the army as police struggled to contain the crowd of thousands outside the US embassy in Islamabad with tear gas and live rounds.
Some protesters had said they would not leave the diplomatic enclave until the US embassy was on fire.
Streets leading to the enclave, where most of the embassies are housed, were earlier blocked off with shipping containers in an effort to increase security.
Television pictures showed chaotic scenes as police tried to gain control of the situation.
Protesters burned an effigy of President Obama and threw missiles at the police.
The BBC's Aleem Maqbool in Islamabad, who did not see any evidence of the army at the scene, said the protest was "turned out like a light".
He said it was amazing, given the strength of feeling at the the protest earlier, that the crowd left as peacefully as it did.
The US State Department earlier issued a warning against any non-essential travel to Pakistan.
It also "strongly urged" US citizens in Pakistan to avoid protests and large gatherings.
Anti-US sentiment has been growing since people became aware of the amateur film earlier this month.
The US Ambassador to Libya was killed in an attack on the US consulate in Benghazi, on 11 September.
Protests in countries around the world have since taken place, with tensions further inflamed by the publication by a French magazine of obscene cartoons of the Prophet Muhammad on Wednesday.
The Pakistani government has called a national holiday on Friday to enable people to demonstrate peacefully.
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