Turkish artillery fired 50 to 60 shells from howitzers positioned in its southern Kilis region against IS targets in the north of Syria's Aleppo province, the private Dogan news agency reported.
A fragile ceasefire has taken effect in Syria, but jihadists are excluded from it.
It was the first attack in several weeks since Turkey, a member of the international coalition against the IS group, stuck jihadist positions in Syria.
Turkish artillery have fired on IS targets in Syria and in Iraq after a deadly suicide attack in Istanbul's tourist hub of Sultanahmet in January.
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The ceasefire deal in Syria, brokered by Russia and the United States, has been in place since Friday midnight. UN chief Ban Ki-moon said there been some incidents but the ceasefire was generally holding.
The deal does not apply to territory held by the IS group and Al-Qaeda affiliate Al-Nusra Front.
Turkish artillery has fired Kurdish militia forces this month, saying it was responding to incoming fire.
Ankara said the Syria truce should have excluded the People's Protection Units (YPG) militia, which it considers to be a terror group linked to its own Kurdish militants.
However the United States works closely with the YPG as the best fighting force on the ground in northern Syria the battle against IS.
The issue has caused major tensions between the two NATO allies, with Washington wanting Ankara to focus on the fight against IS jihadists rather than the Kurdish fighters.