Heineken said it had “consulted with its majority shareholder” before rejecting the approach.
It said the Heineken family, the founding family which still owns half of the firm, wanted to preserve the firm as “an independent company”.
Heineken said it was confident it would continue to grow.
“The Heineken family and Heineken N.V.’s management are confident that the company will continue to deliver growth and shareholder value,” it added.
The founding Heineken family owns just over 50% of the brewer via Heineken Holding. A further 12% is owned by Mexico’s FEMSA.
Heineken’s statement was prompted by a Bloomberg story suggesting SABMiller had approached the Dutch firm to protect itself from a potential takeover.
There has been speculation within the brewing industry, for months, that SABMiller has been targeted by the world’s number one brewer Anheuser-Busch InBev.
Heineken is the world’s third largest beer producer behind AB Inbev and SABMiller, based on volume.
“Should AB Inbev choose to make a bid, which we think is likely, the chance of success has increased,” said Eddy Hargreaves, an analyst at Canaccord Genuity in London.
“SABMiller’s approach may cause some short term share price excitement, but we think Heineken’s decision is final,” he said.
SABMiller gained 5.3% to £35.855 per share in London, while Heineken rose 1.5%.
Apart from the namesake beer – Heineken, the company also produces Sol and Strongbow Apple ciders.
When contacted by the BBC, SABMiller declined to comment. The company is the second-largest beer maker in the world, selling about 21 billion litres of lager worldwide.
SABMiller’s beer brands include Miller Genuine Draft, Grolsch and Peroni Nastro Azzurro. The company also produces soft drinks, and it is one of the world’s largest bottlers of Coca-Cola drinks.
For the full financial year to March 2014, SABMiller earned $27bn (£17bn) in revenues.
SABMiller’s shares are traded on the London stock exchange. The company also has a secondary listing on the Johannesburg stock exchange.