The 20-year-old from the North of France had never been to the fourth round of a Grand Slam before this tournament, but she continued her fine form to set up a meeting with Roberta Vinci.
It looked like Makarova would dominate for much of the opener as both women struggled with their serves early on, and the Russian twice earned leads with breaks before being pegged back by her resilient opponent.
Mladenovic raced into a 3-0 lead in the tie-break, and despite offering a mini-break back, she cruised to a 7-2 success to claim the advantage.
The first nine games of the second set passed by without a break point opportunity as the rivals improved on serve, before Makarova made the key breakthrough to break to love in the 10th game and level the match.
Mladenovic's response was swift and emphatic as she recorded two breaks to win the opening four games of the decider as Makarova started to lose the longer rallies at a pivotal stage of the match.
There was no way back for Makarova as Mladenovic continued to produce winners on a regular basis, and she finished in style with another break to send the 13th seed home.