The rockhole, Anna's Reservoir, was of critical importance to the explorer John McDouall Stuart, who discovered, described and named it in April 1860. He visited it on the forward and return journeys of each of his three attempts to reach the north coast. On Stuart's return from his last, and sucessful, trip in September 1862, the availability of water and grass at Anna's Reservoir at a time when Stuart himself was critically ill and the surrounding area dry, enabled the successful completion of the journey.

The Reservoir once again played an important part in the epic overland journey from 1879-1880 by Alred Giles, overlanding 8000 sheep and 4000 cattle to establish Springvale near Katherine. Giles took advantage of Stuart's description of the water at Anna's Reservoir and forced the sheep over a dry stage of 108 miles from Colyer's Creek (north of Alice Springs) to the Reservoir. This was the only reliable water source in that dry year between Colyer Creek and Tea Tree Well.

The homestead ruins within the Reserve are also highly significant due to their assocation with the Barrow Creek Pastoral Company venture, the most ambitious enterprise of its kind in Central Australia. Billy Benstead, manager of the venture chose in early 1884 Anna's Reservoir as the site of the station homestead. This homestead was the venue for one of the few documented cases where Aboriginal resistance actually forced a withdrawal of white settlement. As such the ruins are important as evidence of that settlement process.

Anna's Reservoir is also important as a wildlife watering hole.