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The Labour party will not do a deal with the SNP "under any circumstances", Sir Keir Starmer will tell party members in Scotland.
In a speech to the Scottish Labour party conference in Edinburgh, he will also warn against complacency following the resignation of Nicola Sturgeon.
And he will appeal to those who have "given up on Labour" and the union.
In response, the Scottish Conservatives pointed to a Labour-SNP deal at Dumfries and Galloway Council.
Until recently Labour was part of a "rainbow alliance" leading the council, despite the party making similar pledges on deals with the SNP before the local elections in May 2022.
Meanwhile the SNP's Mhairi Black said Labour had become "indistinguishable from the Tories on a whole range of issues".
Sir Keir's conference speech comes four days after Nicola Sturgeon resigned as first minister.
Her decision has led to questions about whether a change in SNP leadership could lead to a revival of the Labour party's fortunes in Scotland.
Scottish Health Secretary Humza Yousaf and former minister Ash Regan are the first the announce they will stand to replace Ms Sturgeon.
However Sir Keir will warn party colleagues against complacency.
"We won't change any hearts or minds by sitting back and watching a battle for power within the SNP," he will say.
"We have to go out there and earn it. Prove that Labour can be the change that Scotland needs.
"It's not about change at the top of other parties, it's about the changes we've made - from top to bottom - to our own, and the change we can now offer the Scottish people."
He will say the party has had to show that support for NATO was "non-negotiable", they understand the importance of "sound money" and it "will rip anti-Semitism out by its roots".
The party leader will also repeat a pledge that Labour will not do any deals with the SNP in the event of a hung parliament.
Anas Sarwar, the leader of the Scottish Labour party, made a similar commitment in July last year.
It is an attempt to head off any repeat of the claims by the Conservatives ahead of the 2019 general election that a Labour-SNP pact could lead to another independence referendum.
Sir Keir will tell conference: "Whatever happens in the coming months my message is the same: no deal under any circumstances. The phony offers of support can end now. The blame game can end now.
"The unspoken political bond between the SNP and the Tories, the shared investment in division, that ends now."
However the pledge was criticised by Megan Gallacher, the deputy leader of the Scottish Conservatives, who claimed Labour "can't be trusted to stand up to the SNP".
"Labour made the same pledge before last year's local elections, only for their councillors in Dumfries and Galloway to strike a backroom deal with the nationalists to lock out the biggest party, the Scottish Conservatives."
"Labour can't be the change Scotland needs because you can barely fit a cigarette paper between them and the SNP on most issues," she added.
Power shift
Sir Keir will also confirm his commitment to the union, saying "sticking up a border" will not solve the cost of living crisis, climate change or the response to Putin's invasion of Ukraine.
"But we must accept that many Scots look at Tory Britain and conclude the way out is the way forward," he will add.
He said the Labour party could offer hope to people who had previously "given up on Labour [and]...given up on Britain".
Sir Keir will also touch on devolving more power to the Scottish Parliament as part of a "huge power shift out of Westminster [to] to transform our economy, our politics and our democracy".
Mhairi Black, the SNP's deputy Westminster leader, compared Labour to the Conservatives on the issues of Brexit and Scottish independence.
"This Scottish Labour conference is an opportunity for Starmer to show that he supports the basic principles of democracy and Scotland's right to choose its own future - but we won't hold our breath," she said.
"Similarly, he should recognise that Scotland was forced out of the EU against our will by a Westminster government we didn't vote for - but all we can expect is Starmer doubling down on his hard Brexit rhetoric to win Tory votes in England."