Clare House at Oakville, better known as Wandin Valley Hospital in TV show A Country Practice, is up for auction next month.
The agents said it was only the eighth time in almost two centuries it has been on the market.
The house was a location shoot for the long-running Channel 7 series A Country Practice, from 1981-1993, which at its height was said to have been watched by half Australia’s population.
Several buildings around Pitt Town including the Bird in Hand were used in filming, Molly’s house was at Maraylya, Windsor High was the local high school and the Country Practice clinic was in North Street, Windsor.
Last year what was the Wandin Valley police station in Johnston Street, Pitt Town was sold too.
When the house sold a decade ago, it had been owned by the O’Briens for nearly 30 years.
Gazette journalist Shannon Tonkin talked to Ron and Trish O’Brien at the time. They said a man appeared on their doorstep in 1981 and said he was doing a story about nursing in the bush.
"In less than six weeks, we had film crews here," Mrs O'Brien said.
"They only ever took footage of the outside of our house, but we had our fair share of actors and actresses on the property. They'd be here all day filming for hours, and when you saw it on TV there'd only be five or six seconds of our house, but it was great to see it on the screen."
Mrs O'Brien said many visitors wanted to see 'inside the Wandin Valley Hospital'.
"Contrary to popular belief, it never was a hospital, it's always been a home, but we'd have plenty of people knocking at the door asking to see inside," she said.
The actor who played the show’s character Brendan Jones, Shane Withington, came to Thompson Square in Windsor in December last year to support CAWB’s fight against the new Windsor Bridge plans.
Clare House was built of convict brick on Andrew Thompson’s estate after whom Thompson Square is named, around 1828.
Now on one hectare of land, it also has a dressage arena and new stables with wash bay and kitchen and bathroom facilities. It’s listed with Hamish Rogers.
It will be auctioned on site at 11am on Saturday, September 30, unless sold before.