The BBC's David Stern said it was the second building in the city to be taken over
Pro-Russia activists have seized several official buildings in the eastern Ukrainian city of Luhansk.
The prosecutor's office and the regional administration's headquarters were stormed by armed men.
Interim President Olexander Turchynov criticised local police for their "inaction" and "criminal treachery".
Earlier, Russia criticised sanctions imposed by the US and EU on individuals and companies over their alleged actions aimed at destabilising Ukraine.
Deputy Foreign Minister Sergei Ryabkov said the US had "essentially lowered an 'Iron Curtain'" by targeting Russia's high-tech sector.
The EU, he added, had proved that it was "under Washington's thumb".
Russia's Deputy Prime Minister Dmitry Rogozin meanwhile warned that if the sanctions affected its rocket-building sector, US astronauts on the International Space Station (ISS) might be "exposed".
In other developments on Tuesday:
- An international conference in London heard allegations that Ukraine's ousted President Viktor Yanukovych and his associates may have stolen assets worth tens of billions of dollars
- Ukraine's Interior Minister Arsen Avakov told BBC Russian that voting in next month's presidential election may not be able to take place in all regions because of the unrest in the country's east
- The German government distanced itself from former Chancellor Gerhard Schroeder after he was photographed hugging Russian President Vladimir Putin at a party in St Petersburg
- Norway's defence ministry said it had suspended all defence co-operation with Russia except on border controls with its neighbour
'No control'
Mr Ryabkov also stressed that Russia had no intention of invading eastern Ukraine, where pro-Russia activists have seized government buildings in more than a dozen towns and cities.
Until now, only the local office of the State Security Service (SBU) in Luhansk, a city of 465,000 people less than 30km (20 miles) from the Russian border, had been targeted.
But on Tuesday afternoon, hundreds of people gathered outside the headquarters of the regional government to demand a referendum on granting greater autonomy to the east.
A group of men armed with sticks and metal bars broke into the building, whose entrances were not protected by police. They then pulled down the Ukrainian flag flying from the roof and replaced it with a Russian one, and opened the main entrance to the crowd.
Inside the building's courtyard, they found security personnel in riot gear massed in a defensive position. There was a stand-off, but no violence.
Later, pro-Russia activists stormed the local prosecutor's office and also went into a regional television station, but decided not to take it over after they were allowed to make a live broadcast.
Sanctions
Eastern Ukraine, which has a large Russian-speaking population, was a stronghold for former President Yanukovych before he was overthrown by protesters in February.
The interim government has rejected the pro-Russian activists' demands for greater autonomy, fearing they could lead to the break-up of the country or more regions being annexed by Russia, as happened with Crimea last month.
Pro-Russian activists continue to detain some 40 people, including seven military observers linked to the Organisation for Security and Co-operation in Europe (OSCE) seized last week.
The self-styled "mayor" of the town of Sloviansk, where the observers are being held, has said he will discuss their release only if the EU drops sanctions against separatist leaders.
On Tuesday, the EU published a fresh list of 15 individuals facing travel banks and asset freezes.
Sloviansk resident: "I think that it should end in a peaceful way"
It included Gen Valery Gerasimov, chief of the Russian General Staff, and Lt Gen Igor Sergun, identified as the head of the Russian military intelligence agency, the GRU.
Russian Deputy Prime Minister Dmitry Kozak and pro-Russian separatist leaders in Crimea and in Luhansk and Donetsk were also named.
On Monday, the US announced sanctions against seven individuals and 17 companies it said were linked to President Putin's "inner circle".
The US and EU first imposed visa bans and asset freezes on a number of senior Russian officials and companies after Crimea was annexed.
US Treasury Secretary Jack Lew said the sanctions had so far caused "a quite substantial deterioration in Russia's already weak economy".
Analysis
The pro-Russian gunmen in Ukraine's east seem to be following a strategy of constant expansion and pressure on the Kiev government.
Hardly a day goes by without another incident. Just recently, official buildings in Kostyantynivka have been taken over, Western military monitors detained, peaceful demonstrators in Donetsk attacked, and now the regional administration building in Luhansk has been seized.
It is difficult to say what their ultimate goal is. Perhaps it is to keep government officials in Kiev on the defensive, forcing them to put out a number of fires at once, while others pop up throughout the region.
Or else it is simply to keep the situation unstable, in order to prevent the presidential election scheduled to take place on 25 May.
Or it could be just the opposite, as many in Kiev and throughout the country fear: to provoke the Ukrainians into a full crackdown, which would in turn spark a Russian invasion. The militants have called on Moscow to intervene on more than one occasion.
The most significant effect of the sanctions so far has been on business confidence”
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