The narrator, Sniffer, introduces the Air Buddies, Rosebud, Buddha, Budderball, Mudbud, and B-Dawg and explains their sport talents and personalities. After that, Noah and Henry and their parents leave to go to a basketball game with Buddy and Molly. The pups refuse to be left alone and bored, grab their toys, only to be blocked by their babysitter, Mrs. Niggles and get baths instead. Afterwards, Mrs. Niggles tells them to take naps. B-Dawg decrees that no one shall tell him to take a nap, and leads the pups out to watch the babysitter. Buddha notices the babysitter is 'finding her inner peace' and as she naps, the pups play their favorite sports with their toys and Mrs. Niggles's yarn. After all the victory, Budderball climbs up the counter to claim his prize; Mrs. Niggles's blueberry pie. After throwing it onto the ground, Mrs. Niggles wakes up, but she's tied up by her yarn. Then the pups hear their parents coming home, and because everything is a mess, Mrs. Framm thinks it's time to find the Buddies new homes.
Meanwhile, Selkirk Tander tries to impress Mr. Livingston's son, Bartleby Livingston by showing him a female tiger for his birthday, but Bartleby wanted an animal he can play with. He wants Air Bud (Buddy) because Buddy can play sports. Mr. Livingston offers $500,000 if Selkirk can get Buddy. Selkirk then sends his nephew Grim and assistant Denning to capture Buddy.
At school, Noah and Henry collect every kid's profile sheets and photos, and after looking at girls and boys who would make good owners, the family decides to call the new owners tomorrow. Grim and Denning arrive at Buddy's home, only to see Buddy and Molly at the same time, thinking there are two air buds or they are mates. The next morning, the pups decide to run away. Grim and Denning follow them, and manage to catch Budderball by luring him with a doughnut. The other Buddies go and find Budderball but they are captured as well and used as bait to catch Buddy and Molly. Buddy manages to free the Buddies but Denning traps him with a net. Molly tries to save Buddy but is captured too. The Buddies ask Sniffer for his help and where Buddy and Molly were taken to. Sniffer explains that Buddy and Molly were taken to wine country, and refuses to help because he has lost his sense of smell.
In wine country, Selkirk is pleased that Denning and Grim captured both Buddy and Molly so he can have them breed puppies and sell them for money. When Grim explain that Buddy and Molly already had puppies and Denning let them get away, Selkirk tells them to lock the dogs in the wine cellar and go and capture the puppies and threatens that if they don't catch the puppies, they will be fed to the tiger. Grim and Denning go back to Buddy's home to capture the Buddies but no luck. The Buddies have sniffed their way to the outside cinema, where Grim and Denning are watching 101 Dalmatians. The Buddies find their way to the projection room and walk right in front of the projector, their shadows attracting Denning and Grim's attention. The Buddies then find a snack bar and start eating. Grim opens the projector, temporarily blinding himself. A female customer orders a box cup of popcorn, and Budderball tumbles out of the box—her scream is heard by Denning and Grim who then slip on a wet floor. Next, the Buddies run into a motor bike gang. Still blind, Grim knocks the motor bikes over, while Denning also collides with the motorcycles, furiously calling the Buddies and the man at the movie projection booth double-crossing bottle-suckers and various other nasty names related to double-crossing. Grim and Denning knocking over the motorcycles with their clumsiness infuriates the gang, who then tie Grim and Denning in front of the cinema screen and begin throwing soda, candy, and popcorn at them.
Grim and Denning catch up to the Buddies, chasing them to a farm. The Buddies meet Billy the Goat and Belinda the Pig, who help the Buddies escape from Grim and Denning. The Buddies lure Grim and Denning into a stable and escape through a small hole as Billy locks Grim and Denning in. The Buddies go through the forest then meet the Wolf who leads them to wine country. A skunk's spray enables Sniffer to smell again, so he calls Noah and Henry and they go off to find the Buddies. Buddy and Molly manage to escape to find the Buddies. Budderball falls into a hole, which Buddy and Molly dug, forcing the Buddies to help. Noah and Henry are led by Sniffer through the same farm that the Buddies went through. Noah and Henry are ecstatic and overjoyed at finding Grim and Denning being held prisoners by Billy the goat, and immediately depart to report them and turn them in to the authorities over Grim and Denning's protests. Bartleby and his father come to collect the dogs but instead finds the Buddies. Bartleby and his father then put the Buddies in a limousine, when Noah, Henry, Buddy and Molly come to rescue the Buddies. Budderball falls into one of the wine containers and gets drunk. Bartleby catches Budderball but is caught by Sniffer. Noah, Henry, and the dogs release barrels towards Selkirk and Mr. Livingston; the barrels break the wine container. Selkirk and Mr. Livingston fall in, as the container cracks open. Washing out of the container, Sheriff Bob arrives to arrest Selkirk, Denning, and Grim. The Buddies apologize to Buddy and Molly; they understand and say they're ready for their new owners. Budderball decides to stay with Bartleby because he needs a friend.
- Slade Pearce - Noah Framm
- Christian Pikes - Henry
- Paul Rae - Denning
- Patrick Cranshaw - Sheriff Bob
- Tyler Guerrero - Bartleby Livingston
- Richard Karn - Dr. Patrick Sullivan
- Trevor Wright - Grim
- Cynthia Stevenson - Mrs. Jackie Framm
- Holmes Osborne - Selkirk Tander
- Jane Carr - Mrs. Niggles
- Reese Schoeppe - Sasha
- Scout - Air Bud
- Parker - Air Bud, due to the demanding schedule, more than one dog plays the part of Air Bud
- Abigail Breslin - RoseBud - The only girl and sister of the Buddies. She always wears a pink bow in her fur (usually her head) to separate herself from her brothers. She is also protective of her brothers and her favorite color is pink.
- Dominic Scott Kay - Buddha - The peaceful Buddhist of the Buddies. He loves to meditate and wears a Buddhist collar made of beads. He is named after a virtuous person, Gautama Buddha.
- Josh Flitter - Budderball - The chubbiest of the Buddies. He enjoys food and wears black ink on his face to look like a football player. In addition to this he also sports a red football shirt. When his paw is pulled, he passes gas.
- Spencer Fox - Mudbud - The dirt-loving dude of the Buddies. He wears a blue handkerchief around his neck.
- Skyler Gisondo - B-Dawg - The hip-hoppin' pup of the Buddies. He loves rap music and wears a silver chain with a 'B' as a pendant around his neck to look cool and hip.
- Tom Everett Scott - Buddy - The father of the Buddies. He's still as fun-loving and athletic as he was before the Buddies were born.
- Molly Shannon - Molly - The Buddies mother with golden locks. She loves her children and Buddy very much and will do anything to protect them. She belongs to Henry.
- Michael Clarke Duncan - The Wolf - The Wolf is a friendly inhabitant of the forest who helps the Buddies get to Wine Country.
- Don Knotts - Sniffer - An old Bloodhound who lost his sense of smell years ago. He later regains it after getting sprayed by a skunk. He is one of the characters in the movie who help the Buddies find their parents.
- Wallace Shawn - Billy - A goat who the Buddies meet on the farm. He helps keep the Buddies out of Grim and Denning's clutches.
- Debra Jo Rupp - Belinda - A kind sow who the Buddies meet on the farm. She speaks Pig Latin.
- B-Dawg is the pup who can play basketball, while Budderball plays football, Rosebud plays soccer, Bud-dha plays baseball, and Mudbud plays volleyball and watches Rally America.
- All the animals can speak in this film. This was the first time speaking dogs have appeared in the Air Bud series.
Patrick Cranshaw died on December 28, 2005, but he had already finished filming all of his scenes for the film. Don Knotts died on February 24, 2006, but he finished all his voice work on it. It is dedicated to them both.
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