The conservative Club for Growth and Heritage Action announced Wednesday that they opposes the Senate deal that raises the debt ceiling and ends the government shutdown.
Both groups urged members to vote down the deal, and said they will punish members who support it on their annual scorecards.
The different views highlight the fight between the conservative groups and the Chamber over strategy and tactics, and underline questions about whether business groups will try to punish Tea Party Republicans over their use of the debt ceiling deadline to win concessions.
In a statement, the Club criticized the Senate deal for not having any real reforms in it.
"There are no significant changes to ObamaCare, nothing on the other major entitlements that are racked with trillions in unfunded liabilities, and no meaningful spending cuts either," the Club for Growth said.
"If this bill passes, Congress will kick the can down the road, yet again."
The Chamber criticized the political brinksmanship played in the fight over the government shutdown and raising the debt ceiling.
"In order to prevent another self-made fiscal crisis from occurring in the near future, the administration and Congress must get serious about addressing these long-term issues through a comprehensive deal that includes tax and entitlement reform," said Bruce Josten, the Chamber's executive vice president.
"Regardless, the House and Senate must pass legislation in the coming hours to reopen the federal government and avoid default. The Chamber will include votes on, or in relation to, such legislation in our annual How They Voted scorecard," Josten wrote.
Many House conservatives are expected to oppose the bill when it comes up for a vote, likely late Wednesday.
Speaker John Boehner (R-Ohio) is expected to rely on House Democrats to pass the bill, but has sought to get a majority of his 232 members on board.
This story was updated at 4:02 p.m.