Kane missed Spurs's defeat to Manchester United on Saturday with a hamstring injury but will lead the line for Mauricio Pochettino's side against the Liga champions.
He is joined in the Spurs line-up by England team-mate Dele Alli, who is making his 2017-18 Champions League debut after serving a three-game suspension following a red card against Gent in the Europa League in March.
Madrid coach Zinedine Zidane has chosen a first-choice side to face Spurs, with injury absentees Keylor Navas, Dani Carvajal and Gareth Bale the only regulars not in the line-up.
The clubs find themselves tied atop Group H with seven points, even boasting identical records of seven goals scored and two conceded from three matches.
Tottenham XI: Lloris; Trippier, Sanchez, Alderweireld, Vertonghen; Davies, Dier, Winks, Eriksen; Dele; Kane
Substitutes: Vorm, Aurier, Rose, Dembele, Sissoko, Son, Llorente
Real Madrid XI: Casilla; Achraf, Ramos, Nacho, Marcelo; Kroos, Casemiro, Modric; Isco, Benzema, Ronaldo
Substitutes: Moja, Vallejo, Theo, Vazquez, Asensio, Mayoral, Ceballos