His father, John Bartholomew Sr, started the cartographical establishment in Edinburgh, and he was trained in the firm.

He was subsequently assistant to the German geographer August Petermann, until 1856 when he took up the management of his father's company.

[1] For this establishment, Bartholomew built up a reputation unsurpassed in Great Britain for the production of the finest cartographical work.

He first showcased his colour contouring system at the Paris Exhibition of 1878; although it was initially met with skepticism, it went on to become a standard cartographical practice.

Among his numerous publications, particularly worthy of note is the series of maps of Great Britain reduced from the Ordnance Survey to scales of half-inch and quarter-inch to 1 mile, with relief shown by contour lines and hypsometric tints.