Prior to Harley-Davidson's purchase of the land from the city, the site was formerly used by the Milwaukee Department of Public Works, Lakeshore Sand Company, and Morton Salt.

[9] On the museum's upper level, a procession of motorcycles is displayed down the center of the main hall, running the length of the building, with galleries on either side.

[1] The galleries relate the company's history from its origins in a 10 × 15 foot (3.048 x 4.572 m) wooden shack to its current status as the top U.S. motorcycle manufacturer, producing more than 330,000 bikes each year.

The glass enclosure sits within a floor-embedded, illuminated outline of the backyard shed the motor company was founded in.

The gallery includes a section of a replica wooden board track, suspended in the air at a 45-degree incline.

[6] The museum's upper floor exhibits also include the Gas Tank Gallery, formerly part of the Harley-Davidson 100th Anniversary Open Road Tour.

The exhibit also features exact replicas of the customized Harley-Davidson bikes ridden by Peter Fonda and Dennis Hopper in the 1969 American movie, "Easy Rider", including Fonda's "Captain America" chopper and Hopper's "Billy Bike".

[6] One motorcycle on display was recovered on a beach in British Columbia after being swept out to sea more than 4,000 miles (6,437.376 km) away in a tsunami that hit Japan in 2011.