It is almost 12 years to the day since the diminutive Argentine netted his first, against Albacete in the Camp Nou, and there could not be a better time to bring up the magic number, particularly in a week that began with his great rival Cristiano Ronaldo netting a record 100th Champions League goal.
Enrique proud of Barcelona despite Champions League exit
Barca, who trail Real by three points having played a game more, are under even more pressure than usual after Wednesday's Champions League elimination by Juventus.
The sight of Messi flat out with blood gushing from his face after a heavy tackle from Miralem Pjanic seemed to sum up his club's recent fortunes.
Barca lacked the hunger, fight or determination to break down a stubborn opponent.
What perhaps was most striking as they meekly exited Europe was that even the mercurial Argentine appeared drained of his superpowers against a resilient backline.
He was bullied, harangued and harried out of the game by the Italians.
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In complete contrast to Ronaldo, who almost single-handily dragged Real Madrid through against Bayern Munich 24 hours earlier, the five-time world player of the year looked dejected as the game drew to its inevitable conclusion before sloping off at the final whistle.
However, former teammate Ronaldinho is backing Messi to return to his best. "It's a wonderful story. He's on the verge of 500 goals for the club and my pass set up the first of them," Ronaldinho said. "He's a great man. It's wonderful to see everything he's achieved - and he still has lots left to give."
The Brazilian's words will be put to the test this weekend as, in the absence of the suspended Neymar, the focus on Messi will be even more intense.
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The Argentine will need to be at his brilliant best if Barca are to overcome Zinedine Zidane's in-form side, who are unbeaten in their last 12.
With just two wins in their past five games and dumped out of the Champions League at the quarter-final stage for the second year running, their season is in danger of fizzling out.
Defeat to Real will leave them with just the Copa del Rey to play for.
Although Messi is La Liga's top scorer this season with 29 goals, he has not netted in his past six appearances against Real, dating back to his 2014 hat-trick at the Bernabeu.
In the same period, the Portuguese has scored three in five Clasicos.
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