He was born in Glasgow, on Charlotte Street, the son of Kelso-born banker Alexander Blackie (d.1846) and Helen Stodart.<ref name=fells>Template:Cite web</ref><ref>Template:Cite book</ref> He was educated at the New Academy and afterwards at the Marischal College, in Aberdeen, where his father was manager of the Commercial Bank.Template:Sfn
After attending classes at Edinburgh University (1825–1826), Blackie spent three years at Aberdeen as a student of theology. In 1829, he went to Germany, and after studying at Göttingen and Berlin (where he came under the influence of Heeren, Müller, Schleiermacher, Neander and Böckh) he accompanied Bunsen to Italy and Rome. The years spent abroad extinguished his former wish to enter the Church, and at his father's desire he gave himself up to the study of law.Template:Sfn
He had already, in 1824, been placed in a lawyer's office, but only remained there six months. By the time he was admitted a member of the Faculty of Advocates (1834) he had acquired a strong love of the classics and a taste for letters in general. A translation of Goethe's Faust, which he published in 1834, met with considerable success, winning the approbation of Carlyle. After a year or two of desultory literary work he was (May 1839) appointed to the newly instituted Chair of Humanity (Latin) in the Marischal College.Template:Sfn
Difficulties arose in the way of his installation, owing to the action of the Presbytery on his refusing to sign unreservedly the Confession of Faith; but these were eventually overcome, and he took up his duties as professor in November 1841. In the following year he married. From the first his professorial lectures were conspicuous for the unconventional enthusiasm with which he endeavoured to revivify the study of the classics; and his growing reputation, added to the attention excited by a translation of Aeschylus which he published in 1850, led to his appointment in 1852 to the Professorship of Greek at Edinburgh University, in succession to George Dunbar, a post which he continued to hold for thirty years.Template:Sfn
He was somewhat erratic in his methods, but his lectures were a triumph of influential personality. A journey to Greece in 1853 prompted his essay On the Living Language of the Greeks, a favorite theme of his, especially in his later years; he adopted for himself a modern Greek pronunciation, and before his death he endowed a travelling scholarship to enable students to learn Greek at Athens.Template:Sfn
During J. S. Blackie's tenure of the Greek Chair, he taught Robert Louis Stevenson, and Stevenson would later write:
'Although I am the holder of a certificate of attendance in the Professor's own hand, I cannot remember to have been present in the Greek class above a dozen time. Professor Blackie was even kind enough to remark (more than once) while in the very act of writing the document above referred to, that he did not know my face.'<ref>Template:Cite book</ref>
Scottish nationality was another source of enthusiasm with him; and in this connection he displayed real sympathy with highland home life and the grievances of the crofters. The foundation of the Celtic chair at Edinburgh University was mainly due to his efforts. He raised almost all £12,000 needed for it.<ref name="John Stuart Blackie">Template:Cite web</ref> In spite of the many calls upon his time he produced a considerable amount of literary work, usually on classical or Scottish subjects, including some poems and songs of no mean order.Template:Sfn
Blackie was a Radical and Scottish nationalist in politics, of a fearlessly independent type; possessed of great conversational powers and general versatility, his picturesque eccentricity made him one of the characters of the Edinburgh of the day, and a well-known figure as he went about in his plaid, worn shepherd-wise, over one shoulder and under the other, wearing a broad-brimmed hat, and carrying a big stick.Template:Sfn He was made the first chairman of the Scottish Home Rule Association.<ref>Template:Cite book</ref>
In the 1880s and 1890s, he lectured at Oxford on the pronunciation of Greek, and corresponded on the subject with William Hardie. In May 1893, he gave his last lecture at Oxford, but afterwards admitted defeat, stating: "It is utterly in vain here to talk reasonably in the matter of Latin or Greek pronunciation: they are case-hardened in ignorance, prejudice and pedantry".<ref>Template:Cite book</ref>
He died at 9 Douglas Crescent<ref name=fells/> in Edinburgh.<ref>Template:Cite news</ref> His death caused great mourning in the city.<ref name="John Stuart Blackie"/> He is buried in Dean Cemetery to the north side of the central path in the north section of the original cemetery. His nephew and biographer, Archibald Stodart Walker (1869-1934) is buried with him.
Before his death John Blackie gave more than 250 volumes of 19th-century Greek books on various subjects to the University of Edinburgh library.<ref name="John Stuart Blackie"/>
In 1895, a plaque, designed by Robert Lorimer was erected to his memory in St Giles Cathedral.<ref>Dictionary of Scottish Architects: Robert Lorimer</ref>
Messis Vitae: Gleanings of Song from a Happy Life. 1886.
The Lyrical Dramas of Æschylus, Translated Into English Verse by John Stuart Blackie, Two Volumes. 1850.
Amongst his political writings, may be mentioned:
On Democracy, Edmonston & Douglas, 1867.<ref>Jones, Ernest Charles. Democracy Vindicated: A Lecture Delivered to the Edinburgh Working Men's Institute on 4 January 1867, in Reply to Professor Blackie's Lecture on Democracy, Delivered on the Previous Evening, Andrew Elliot, 1867.</ref>
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