The Czech Republic international, who is out of contract at the end of the season, has recovered from a hamstring problem.
With Morgan Fox ruled out for the remainder of the campaign through injury, Pudil is poised to partner Tom Lees and Frederico Venancio in Wednesday’s three-man defence.
Boss Jos Luhukay told The Star: “I’m very happy Daniel is available. He was not 100 per cent for the game at Sunderland but he is 100 per cent for Saturday.”
Fox and Joost van Aken have been added to a long list of players who will not figure again this season.
The Owls have climbed up to 16th in the table following a three-match winning run which has all but guaranteed their Championship safety.
“In the time that I have been here, the atmosphere has always been good,” said Luhukay, who could be without Keiren Westwood for the final six matches. “The team have stayed together and that was the same when we were losing games.
“The character and team spirit is very positive.
“Of course, you see only the 90 minutes and the result sometimes puts things in a positive or negative direction. I see the players every day and how they work and how they are growing with each other.
“It has given me a good feeling since the first day and that we can come to success in the next few games. We must also look to the future and it is my job to do that.”