The measure comes following a public consultation which was carried out between August and November.
Estimates suggest that, while 24 million adults in Great Britain gamble, 800,000 consumers use credit cards to gamble.
The Commission’s own research indicates 22% of online gamblers using credit cards to gamble are assessed as problem gamblers.
Neil McArthur, Gambling Commission CEO, said: "Credit card gambling can lead to significant financial harm. The ban we have announced should minimise the risks of harm to consumers from gambling with money they do not have.
"We also know there are examples of consumers who have accumulated tens of thousands of pounds of debt through gambling because of credit card availability.
"There is evidence the fees charged by credit cards can exacerbate the situation because the consumer can try to chase losses to a greater extent."
McArthur did emphasise the Commission’s awareness of consumers who gamble responsibly with the use of credit cards, suggesting the regulator will evaluate the measure for any "unintended circumstances."
The Gambling Commission CEO added: “We realise this change will inconvenience those consumers who use credit cards responsibly but we are satisfied reducing the risk of harm to other consumers means action must be taken.
"But we will evaluate the ban and watch closely for any unintended circumstances for consumers."
The share prices of major UK operators have fallen since reports of the ban first surfaced at the start of the week.
GVC Holdings' share value has dropped from an opening price of £9.42 ($12.24) on Monday to £9.14; Flutter Entertainment's share price has fallen from £94 to £91.94; meanwhile, William Hill's shares saw a similar decrease from £1.94 to £1.83.
