Galatasaray should take confidence from successive away victories against Cluj, SC Braga and Schalke 04 in this season's European campaign but Real present their toughest challenge yet, particularly at the Bernabeu.
Madrid have won 13, and lost one, of 16 European home fixtures since the appointment of Jose Mourinho.
The Madrid boss will field his strongest side on Wednesday, having rested the likes of Xabi Alonso, Mesut Ozil and Angel Di Maria at the weekend.
Raphael Varane and Fabio Coentrao are also expected to return after missing the 1-1 draw with Real Zaragoza with slight muscle injuries.
But goalkeeper Iker Casillas will only be on the bench, despite recovering from a hand injury, meaning Diego Lopez will start in net.
While Real have been eliminated in the semi-finals in the past two seasons, it is almost a quarter of a century since Galatasaray last reached the last four.
However, their cause is boosted by having two Champions League winners among their ranks who Mourinho knows very well.
Striker Didier Drogba won the Premier League twice under Mourinho's management at Chelsea between 2004 until 2007, before the Portuguese departed Stamford Bridge.
Drogba went on to further glory with Chelsea, scoring the winning penalty in last year's final as they beat Bayern Munich in a shoot-out after having also scored the Blues' equaliser in the Allianz Arena.
The 35-year-old quit the London club for a stint in China before moving to Turkey and he will lead the line against Real.
Alongside Drogba in attack will be Dutchman Wesley Sneijder, who won the Champions League with Mourinho at Inter Milan in 2010.
Galatasaray also have a number of other players with Champions League experience such as former Arsenal player, Felipe Melo and Fernando Muslera, who have represented Juventus and Lazio respectively.
The match also pits the two top scorers in this year's tournament against each other, with Burak Yilmaz lining up for the away side and Cristiano Ronaldo in the home ranks. Both have eight goals to their name.