A person familiar with the sale process confirmed to The Associated Press on Thursday that Golisano submitted a non-binding bid to Morgan Stanley, the banking firm overseeing the sale. The person spoke on the condition of anonymity because the sale is private.
The Buffalo News first reported Golisano's entry earlier in the day.
The Bills are on the market after Hall of Fame owner Ralph Wilson died in March.
Golisano's addition comes at the same time the sale process moved into the next phase after Bills officials began meeting with prospective ownership groups that had already submitted their non-binding bids.
Two other people familiar with the process said New York real estate mogul Donald Trump was first up, by attending a lengthy presentation in the Detroit-area on Wednesday.
Current Sabres owners Terry and Kim Pegula, and a Toronto-based group fronted by rocker Jon Bon Jovi are also scheduled to attend presentations.
The presentations provide prospective buyers an opportunity to receive extensive financial and background information on the franchise in order to prepare their formal bids, which are expected to be submitted within three weeks.
A prospective owner is expected to be identified by the end of next month, and then presented to NFL owners for approval at league meetings in New York in early October.
Hall of Fame quarterback Jim Kelly and his brother Dan Kelly, who runs the family's business interests, have also been invited to a presentation. Though the Kellys have expressed interest in purchasing the team, it's unclear if they have submitted a bid or have linked up with another ownership group.
Golisano had first indicated an interest in purchasing the Bills in May. Though Golisano now lives in Florida, he is from western New York, and made his fortune establishing the Rochester-based payroll firm, Paychex.
He purchased the Sabres out of bankruptcy in 2003, before selling the team to Pegula in 2011.
Golisano's net worth was most recently valued by Forbes to be $1.9 billion.
Though prospective buyers had until July 29 to submit their non-binding bids, Morgan Stanley extended the deadline to allow for other groups to enter the running.
The Pegulas and Trump are considered the front-runners. Both have the assets to complete what is expected to be a $1 billion-plus purchase, and they also have expressed a commitment to keep the Bills in western New York.
Terry Pegula was last valued by Forbes to have a net worth of $3.3 billion. That was before Pegula closed a $1.75 billion deal on Tuesday to sell about 75,000 acres of drilling rights to American Energy Partners.
Trump was last valued with a net worth of $3.9 billion. That does not include his holdings in numerous properties, including several high-profile golf resorts he recently purchased.
The Bon Jovi-fronted group is made up of Toronto-based partners, Larry Tanenbaum, chairman of Maple Leaf Sports and Entertainment, and the Rogers family, which controls Rogers Communications.
However, it's unclear whether Bon Jovi has the liquid assets to buy the Bills if the sale price goes above $1.2 billion.
Under NFL rules, a principle owner must have enough cash and use no more than $200 million in debt-financing to hold a minimum 30 per cent ownership stake.
Forbes most recently valued Bon Jovi's net worth at $290 million.
Bon Jovi also faces questions about his long-term intentions to keep the team in the Buffalo area, which is a prerequisite in the sale process.
Last weekend, Bon Jovi had a letter published in The Buffalo News stating that his objective was "to carry on the legacy of Ralph Wilson and make the Bills successful in Buffalo."
He didn't state in the seven-paragraph letter that he was committed to keeping the franchise in Buffalo. The only commitment Bon Jovi provided was working with state and local officials to identify a site for a potential new stadium.
The letter was received with much skepticism in Buffalo and among Bills fans, who fear Bon Jovi eventually intends to relocate the team to Toronto.
Under the current lease, reached in December 2012, the Bills would incur a $400 million penalty for broaching the prospect of moving during the 10-year agreement. There is a one-time exception that would allow the Bills to break the agreement for just under $28.4 million in 2020.
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