Senator Wong will meet regional counterparts during her visit to Israel and the Occupied Palestinian Territories, along with Jordan and the United Arab Emirates.
Aid group Amnesty International said Senator Wong's visit to the region should be focused on ways for a ceasefire to be undertaken.
The organisation's Occupied Palestinian Territories and Israel spokesman Mohamed Duar said the 100 days since the start of the war had seen a "chilling disregard of human rights".
"Amnesty has repeatedly documented war crimes being committed by all parties to the conflict,'' he said.Â
"The scale of death and destruction we witness in Gaza today is unparalleled.
"For these reasons, Foreign Minister Penny Wong must reiterate Australia's support for a ceasefire, and for Israel to allow unhindered and immediate humanitarian aid to enter Gaza during her trip."
Senator Wong will be the most senior representative from Australia to visit the region since the current conflict broke out on October 7 and the first from an Australian foreign minister since 2016.
Before her departure, the foreign minister said it was critical to advocate for a peaceful resolution.
"Australia is not a central player in the Middle East, but we are a respected voice, and I'll be using our voice to advocate for a pathway out of this conflict," she said.
Senator Wong condemned settler violence within the region, while reiterating support for a two-state solution.
"Attacking Palestinians where they have a right to be is the wrong thing to do and certainly not conducive to ensuring there isn't escalation," she said.
Mr Duar said Amnesty welcomed the foreign minister's commitment to address settler violence in the West Bank.
But Senator Wong had come under fire for not visiting the site of massacres following attacks by Hamas, which has been listed as a terrorist organisation by the federal government.
Save the Children's country director for the Occupied Palestinian Territory Jason Lee said a ceasefire was needed in the region.
"For every day without a definitive ceasefire, 100 children on average have been killed," he said.
"There can never be any justification for killing children."
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