Scottish Labour leader Richard Leonard has said Labour will commit to opposing a second independence referendum in its next UK manifesto.
He believes this would give Jeremy Corbyn a mandate to refuse permission for such a vote to be held.
First Minister Nicola Sturgeon tweeted that Labour is likely to alienate its own former supporters with this policy.
Earlier this week, Mr Corbyn told BBC Scotland he would "decide at the time" if he would approve a section 30 order.
This is needed for the Scottish government to lawfully hold a referendum.
Speaking on the BBC's Sunday Politics Scotland programme, Mr Leonard said: "We want to be absolutely clear to the people of Scotland that there is no case for a second independence referendum.
"We just had a referendum in 2014. We think that settled the will of the people of Scotland.
"I don't think there is any support for a second independence referendum, which is why we are going to categorically - in our manifesto in the lead up to the general election which may come as soon as later this year - state our opposition to the holding of a second independence referendum."
Mr Leonard's comments have been strongly criticised by supporters of Scottish independence.
In a tweet shortly after the interview was broadcast, Ms Sturgeon said: "Scottish Labour's determination to remain alienated from swathes of its previous support as it attempts to out Tory the Tories on #Indy shows no sign of abating. Which can only be good news for @theSNP"
SNP deputy leader Keith Brown later told BBC Scotland: "I think they display an extraordinary contempt for the democratic rights of the people of Scotland.
"In the same way that we've seen it over Brexit, with Scotland being pulled out of the EU against its will.
"What we're now seeing is Richard Leonard putting the Labour party in a position where even when there's a democratic mandate for Scots to choose in the future for independence, he would oppose that."
Labour leader Jeremy Corbyn said on Thursday he did not support another referendum but was "not ruling out" giving consent for a vote if he were to become prime minister.
Asked by BBC Scotland what he would do if First Minister Nicola Sturgeon were to seek the power to hold another vote, he said: "We would obviously decide at the time."
He added: "We don't want another referendum, we don't think another referendum is a good idea, and we'll be very clear on why we don't think it's a good idea."