Angela Rayner has urged Sir Keir Starmer to stand firm and deliver her workers' rights reforms.
The former deputy prime minister said the Employment Rights Bill, which she championed in office, was "good for workers and it is good for business".
The Tories claim it will cost businesses billions, and even the Left-wing Resolution Foundation has warned that the new right to claim unfair dismissal on day one will cost jobs.
Speaking in the Commons, Ms Rayner said it was the Government's duty to overturn the changes and deliver its manifesto promises.
It is her first major intervention since she was forced to resign from her ministerial positions after The Telegraph exposed details of her tax affairs.
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"This Bill was a promise we made to the British public," she said. "It is our duty to deliver it. And I say to my colleagues on the front bench that I will be with you every step of the way as we do just that.
"Make no mistakes. The bill is good for workers and it is good for business. This isn't just the right thing to do, it's the foundation for the high-growth, high-skill economy that the UK needs."
She added: "We believe workers deserve fairness, dignity and respect at work, and they deserve it from day one on the job."
It comes amid rumours that the Prime Minister is considering watering down aspects of the bill over fears it will hamper growth.
Last month the House of Lords defeated the Government and changed the bill to bring in a qualifying period of six months before unfair dismissal protections come in.
But on Wednesday, the Government rejected the Lords' changes. Kate Dearden, the business minister, reiterated that the Government was "committed to delivering unfair dismissal protections from day one".
"Day one protection from unfair dismissal will not remove the right of businesses to dismiss people who cannot do their job or pass a probation, but it will tackle cases of unfair dismissal, where hard-working employees are sacked without a good reason," she said.
"A six-month qualifying period threshold still leaves employees exposed to dismissal without good reason in the early months of a new job."
She also rejected a Lords proposal to change the bill's provisions over guaranteed hours.
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Under their amendment, employers would have to notify workers of their right to guaranteed hours and make an offer of guaranteed hours, unless the workers decline or opt out.
Ms Dearden said this would "undermine the Bill's core aim of ending exploitative contracts and provide security for the workers who need it most".
Shadow business secretary Andrew Griffith branded the bill a "clunking fist of regulation dictating and providing perverse incentives, maybe unintended consequences, that mean employers simply won't take a chance on those young people at all".
He said it would risk hurting British businesses and make it harder for "vulnerable" workers such as young people, disabled people, those over 50, and former prisoners to get a job.
Noting the word "union" appears 478 times in the bill, and the number of Labour MPs who receive donations from unions, Mr Griffith urged the Commons to "follow the money".
A Lords bid to keep the 50 per cent turnout threshold for industrial action ballots was also rejected by the government on Wednesday.
Ms Dearden said scrapping the threshold "removes an unnecessary bureaucratic hurdle and aligns union democracy with other democratic processes, such as parliamentary votes and local elections, which do not typically require turnout thresholds, but are still accepted as legitimate".
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