A rested Balotelli is set to lead Milan's attack alongside Alessandro Matri. The 23-year-old striker, who is Milan's leading scorer this season with four goals, returns after missing his team's last two Serie A games through suspension.
Balotelli was punished by the Italian league for insulting a referee during a 2-1 defeat to Napoli.
"In the past few days I've had a knee problem but I am ready to play against Ajax," Balotelli told Gazzetta dello Sport.
"In Amsterdam, we want to give continuity to our victory over Celtic. Our aim is to arrive as calmly as possible to our double-header with Barcelona."
Milan won 2-0 against Celtic in their group opener and go into their midweek encounter inspired by Saturday's 1-0 triumph over Sampdoria, courtesy of Valter Birsa's goal.
The result, Milan's second league victory this season, halted a three-match winless run and lifted the Rossoneri to ninth place.
"We have been unlucky at the start of the season," Balotelli said.
"But we are not inferior to any team and I'm sure that by Christmas we will be in the top positions."
Milan have travelled to Holland without five injured players but the good news for coach Massimiliano Allegri is that midfielder Riccardo Montolivo and striker Stephan El Shaarawy have returned to fitness and could feature against Ajax.
Ajax, meanwhile, endured a dismal start to the group stages with a 4-0 defeat at Barcelona earlier this month.
The Dutch giants, whose last victory against a Serie A team in European competition was a 2-1 triumph over Roma 11 years ago, warmed up for Tuesday's game by demolishing Go Ahead Eagles 6-0 at the weekend.
The victory lifted Ajax to fifth position in the Eredivisie.
Ajax coach Frank de Boer was impressed by his team's display and expects more of the same when his side take on Milan.
"I wanted to see an Ajax that was aggressive in play, coaching, and fending off the opponent," De Boer said.
"And I saw that (against Go Ahead Eagles) for the entire 90 minutes. Then you see that both attacking and defending can be beautiful.
"We controlled and created opportunities. We showed passion and that is what we need to do on Tuesday."
The hosts will be without forward Bojan Krkic, who sustained a hamstring injury on Saturday.
Milan striker El Shaarawy, who has already missed a month of action through injury, was later ruled out of the trip after suffering a hairline fracture to a metatarsal bone in his left foot during a final training session at Milanello.
Vice-president Adriano Galliani told the club's website: "It's a real shame but we'll get him back soon."