Health officials in Gaza said the Sunday night strike killed at least 45 people in a displaced persons' camp in Rafah, the south Gaza city where Israel launched a controversial offensive earlier this month.
Sadr condemned the Israeli strike and Washington's "shameless" support for the "genocide" he charged was under way in Gaza.
"I reiterate my demand to expel" the US ambassador and "close the embassy through diplomatic means without bloodshed," he said in a statement on X.
He said that would be a more effective deterrent than the use of force and would mean US officials "don't have an excuse to destabilise Iraq".
Sadr once led a militia fighting US-led forces after the 2003 invasion that toppled longtime dictator Saddam Hussein.
He retains a devoted following of millions among the country's Shiite Muslim majority community, and wields great influence over Iraqi politics.
The Iraqi foreign ministry condemned the "criminal acts that the occupation continues to commit" in Gaza, and urged the international community to take "deterrent" steps and impose sanctions on Israel.
The Israeli strike prompted a wave of international condemnation.
Israeli Prime Minister Benjamin Netanyahu said it was a "tragic accident" but vowed to push on with the military campaign to destroy Hamas.
The war in Gaza began when Hamas militants attacked Israel on October 7, resulting in the death of around 1,170 people, mostly civilians, according to an AFP tally of Israeli official figures.
Israel's retaliatory offensive has killed at least 36,050 people in Gaza, mostly civilians, according to the Hamas-run territory's health ministry.
All of Iraq's political parties support the Palestinian cause. Like its neighbour Iran, Iraq does not recognise the Israeli state.
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