The "slow and bumpy" 4X4, built in 1950, featured in a scene with soap opera star Nick Berry, who played PC Nick Rowan in the period drama set in Yorkshire.
Heartbeat was a popular ITV drama - based on the 'Constable' books by Nicholas Rhea - which ran for 18 seasons from 1992 to 2010.
The Land Rover, which is still in working order with its original 1600cc engine, featured in series 2, episode 5, which aired in 1993.
Now, a classic car collector is parting ways with the Series 1 Land Rover as it's set to sell on October 23 at Cheffins auction house, Cambs.
Bill King, Chairman at Cheffins says: “Whilst they are slow, bumpy and hugely uncomfortable, the Land Rover Series are the original and the best of the 4x4.
"As the ultimate British vehicle, no other model or make will come anywhere near the Land Rover’s eponymous design and style. F
"From agricultural workhorse to the modern-day countryman’s Ferrari, these series versions were originally built to plough the fields like a tractor, but are now seen as hugely glamorous and sought-after."
Mr King added that there could be a rise in demand for Series Land Rovers as one features in the new James Bond film, No Time to Die.
He added: "We expect that following the Series III seeing a starring role in the upcoming No Time To Die film that Series Land Rovers will become even more popular, especially with those who fancy themselves as a bit of a 007.
"There are two different markets here, some buyers look for fully-restored tidy examples, whilst there are also the collectors out there who are keen for a project.”
Cheffins estimates that the Land Rover will sell for £14,000-16,000.