Matthew Corbett runs a shop in Manchester, with puppets Sooty (a yellow bear that can perform magic), Sweep (a squeaking dog), Soo (a female panda), and "little cousin Scampi" (another, if smaller, bear). Sooty and the gang are often getting up to mischief, and it is usually Soo and Matthew that are the more sensible ones.
A regular visitor at Sooty and Co., is Mo (from the market), and most episodes involve Matthew getting into a mess due to the puppets, whether it is having water poured over him or having custard pies thrown at his face. Some episodes finish with a song, usually performed by Matthew and the group.
- Matthew Corbett the show's presenter.
- Sooty - a mute yellow bear who is the protagonist of the show. He owns a magic wand whose power is invoked using the words "Izzy wizzy, let's get busy".
- Sweep - a dim-witted grey dog with a penchant for bones and sausages. He wears red wellingtons and communicates using bizarre squeaks.
- Soo - a calm and collected female panda who acts as the foil for both Sooty and Sweep. Usually wears a red skirt. Voiced by Brenda Longman.
- Little Cousin Scampi - Sooty's cousin, who is very cheeky and a serial troublemaker. He wears a navy blue school uniform and, like Sooty, is mute.
- Mo from t'market - a regular customer with a blonde perm. Always introduces herself with "Hello, it's only me, Mo from t'market." Played by Brenda Longman.
- Richard - Richard was introduced into the series for the final season, as a new employee that Sooty & Co. had taken on. In reality, he was introduced so the children could get to know the new presenters
- Liana Liana was introduced into the series for the final season, as a new employee that Sooty & Co. had taken on. In reality, she was introduced so the children could get to know the new presenters.
- Unlike "The Sooty Show" where he narrated in the third person, Matthew speaks in first person on this show, which he also did in Learn with Sooty.
- The first episode of Sooty & Co.(along with The Sooty Show) was shown on CITV's Old Skool Weekend to celebrate 30 years of Childrens ITV.