Lawmakers in the Edinburgh legislature passed a motion Tuesday refusing to consent to the EU Withdrawal Bill.
Prime Minister Theresa May is under no obligation to amend her Brexit plan if Scotland objects, but experts warn that a confrontation between London and Edinburgh could push Scotland towards independence.
An initial proposal past year by Britain that devolved powers returning from the European Union after Brexit should initially pass to Westminster was roundly rejected by Welsh and Scottish politicians.
Scotland voted against independence by 55 percent in a referendum in 2014, but Sturgeon insists she has a mandate to hold a second vote since Scotland voted against Brexit by 62 percent in 2016.
Speaking after the vote, he said any move to force legislation on Holyrood would break a "20 year old devolution settlement". The Scottish National Party joined forces with Labour, the Liberal Democrats and the Greens to vote down Theresa May's European Union withdrawal bill.
"The Scottish Parliament has spoken loudly and clearly - it's now up to the United Kingdom government to respect that vote and ditch their power-grab", SNP Scottish lawmaker Ash Denham said in a statement.More news: This cup has to be mine, says Neymar
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Meanwhile, the UK Government has accused Scottish opponents of "nit-picking" and told its Scottish counterparts the "door is still open" for a deal to end the long-running Brexit powers dispute.
It has never been done before by the devolved Parliament in Holyrood.
"Obviously, there'll be an opportunity for further debate and discussion in parliament but also I hope there'll be the opportunity for debate and discussion between the two governments".
The British government said it would refer that bill to legal officers and a hearing is scheduled for July in the Supreme Court unless agreement can be found before then. Ministers in both governments are discussing a fresh round of talks.
Nicola Sturgeon has suggested the Scottish Parliament vote could be "overturned" if a deal is reached.
The Scottish Government argues that all powers not now reserved to Westminster should automatically be devolved.
The Scottish vote came as Ms May's cabinet subcommittee on Brexit met amid enduring differences over Britain's customs relationship with the European Union after Brexit.