The foreign minister leaves on Monday to travel to Israel, Jordan, the Occupied Palestinian Territories and the United Arab Emirates over the week.
Senator Wong will meet with the Israeli families of hostages and survivors of the October 7 attacks.
She will also meet with Palestinians impacted by Israeli settler violence in the West Bank.
Speaking in Adelaide before her departure, the foreign minister deflected a question about why she was not visiting the sites of the massacres perpetrated by Hamas militants, as other political leaders had.
In response to reporters, she said her meeting with the families and survivors of the attacks "will be important".
Sunday marked 100 days since terror group Hamas killed 1200 Israelis and took another 240 hostage.
Israel has bombarded Gaza with air strikes. (EPA PHOTO)
Israel has since bombarded Gaza with air strikes which local authorities say have killed more than 23,000 people and displaced 1.9 million, or 85 per cent of the besieged strip's population.
The trip comes just days after Australia supported the US and UK's bombing of Houthi rebel controlled territory in Yemen following the Iran-backed group's attacks on commercial shipping in the Red Sea.
The rebels have launched assaults in the trade route in response to Israel's strikes on Gaza.
Prime Minister Anthony Albanese rejected a suggestion that Australia's support for the joint action against the Houthis had been inadequate.
"We're making an appropriate contribution as Australia always does," he told ABC's RN.
"We always play our role and it is appropriate that we have people there in Bahrain, we had people before the Houthi attacks.
"Since then we've increased the number of personnel that are involved in that operation."
Senator Wong will balance Australia's unequivocal condemnation of Hamas' attack and support for Israel's right to defend itself, against the path towards a ceasefire.
"Israel must respect international humanitarian law and conduct military operations lawfully. Civilians and civilian infrastructure must be protected," she said.
"I will express our profound concern that there are increasingly few safe places for Gazans.
"I will reiterate our call for safe, unimpeded and sustained humanitarian access so that food, water, fuel, medicine and essential assistance to reach people in desperate need, and so civilians can get to safety.
"Australia wants to see steps towards a sustainable ceasefire."
Senator Wong said a ceasefire could never be "one-sided".
"It is our view that Gaza must no longer be used as a platform for terrorism and that Hamas must lay down its arms," she said.
"We are committed to working with partners toward a just and enduring peace in the form of a two-state solution, where Israelis and Palestinians can live securely within internationally recognised borders."
The government said the visit to Israel - the first by a foreign minister since 2016 - built on Senator Wong's extensive diplomatic efforts since the crisis began.
Senator Wong will meet with her regional counterparts to discuss preventing the spread of conflict and the ongoing humanitarian crisis.
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