Rob Roy was born at Glengyle, near Loch Katrine. His father was Donald MacGregor and his mother Margaret Campbell. In January 1693 Rob married Mary Helen MacGregor of Comar. They had four sons: James (known as Mor or Big), Ranald, Coll, and Robert (known as Robin Oig or Young Rob).
Along with many Highland clansmen, at the age of eighteen Rob Roy together with his father joined the Jacobite rising, to support the StuartKing James II who had been deposed during the Glorious Revolution of 1688. Although victorious in initial battles, Dundee was killed in 1689, deflating the rebellion. Rob's father was taken to jail, where he was held on treason charges for two years. Rob's mother Margaret's health failed during Donald's time in prison. By the time Donald was finally released, his wife was dead. The Gregor chief never returned to his former spirit or health.
Rob Roy was badly wounded at the Battle of Glen Shiel in 1719, in which a British army of Scots and English defeated a Jacobite and Spanish expedition that aimed to restore the Stuart monarchy.
Rob Roy became a well-known and respected cattleman — this was a time when cattle rustling and selling protection against theft was a commonplace means of earning a living. Rob Roy borrowed a large sum to increase his own cattle herd, but due to the disappearance of his chief herder, who was entrusted with the money to bring the cattle back, Rob Roy lost his money and cattle, and defaulted on his loan. As a result, he was branded an outlaw, and his wife and family were evicted from their house at Inversnaid, which was then burned down. After his principal creditor, James Graham, 1st Duke of Montrose seized his lands, Rob Roy waged a private blood feud against the duke until 1722, when he was forced to surrender. Later imprisoned, he was finally pardoned in 1727. He died in his house at Inverlochlarig Beg, Balquhidder, on 28 December 1734.
In 1716 Rob Roy moved to Glen Shira for a short time and lived under the protection of John Campbell, 2nd Duke of Argyll also known as Red John of the Battles, "Iain Ruaidh nan Cath." Argyll negotiated an amnesty and protection for Rob and granted him permission to build a house in the Glen for the surrendering up of weapons. "Traditionally the story goes that Argyll only received a large cache of rusty old weapons." A sporran and dirk handle which belonged to Rob Roy can still be seen at Inveraray Castle. Rob Roy only used this house occasionally for the next three or four years.
In July 1717, Rob Roy and the whole of the Clan Gregor were specifically excluded from the benefits of the Indemnity Act 1717 which had the effect of pardoning all others who took part in the Jacobite rising of 1715.
Some time around 1720 and after the heat of Rob's involvement at the Battle of Glen Shiel had died down, Rob moved to Monachyle Tuarach by Loch Doine and some time before 1722 Rob finally moved to Inverlochlarig Beag on the Braes of Balquhidder.