In 1888, Nobel was astonished to read his own obituary, titled The merchant of death is dead, in a French newspaper. It was Alfred's brother Ludvig who had died; the obituary was eight years premature. The article disconcerted Nobel and made him apprehensive about how he would be remembered. This inspired him to change his will.

Signing his well at the Swedish–Norwegian Club in Paris on 27 November 1895. To widespread astonishment, Nobel's last will specified that his fortune be used to create a series of prizes for those who confer the "greatest benefit on mankind" in physics, chemistry, physiology or medicine, literature, and peace. Nobel bequeathed 94% of his total assets, 31 million SEK (US$186 million in 2008), to establish the five Nobel Prizes.

On 10 December 1896, Alfred Nobel died in his villa in San Remo, Italy, from a cerebral haemorrhage. He was 63 years old.

Owing to skepticism surrounding the will, it was not approved by the Storting in Norway until 26 April 1897.

Nobel's instructions named a Norwegian Nobel Committee to award the Peace Prize, the members of whom were appointed shortly after the will was approved in April 1897. Soon thereafter, the other prize-awarding organizations were designated. These were Karolinska Institute on 7 June, the Swedish Academy on 9 June, and the Royal Swedish Academy of Sciences on 11 June.

The executors of the will, Ragnar Sohlman and Rudolf Lilljequist, formed the Nobel Foundation to take care of the fortune and to organise the awarding of prizes.the Nobel Foundation's newly created statutes were promulgated by King Oscar II.

The Curie family has received the most prizes, with four prizes awarded to five individual laureates. Marie Curie received the prizes in Physics (in 1903) and Chemistry (in 1911). Her husband, Pierre Curie, shared the 1903 Physics prize with her. Their daughter, Irène Joliot-Curie, received the Chemistry Prize in 1935 together with her husband Frédéric Joliot-Curie. In addition, the husband of Marie Curie's second daughter, Henry Labouisse, was the director of UNICEF when he accepted the Nobel Peace Prize in 1965 on that organisation's behalf.

In 1938 and 1939, Adolf Hitler's Third Reich forbade three laureates from Germany (Richard Kuhn, Adolf Friedrich Johann Butenandt, and Gerhard Domagk) from accepting their prizes. Each man was later able to receive the diploma and medal. Even though Sweden was officially neutral during the Second World War, the prizes were awarded irregularly.

No prize was awarded in any category from 1940 to 1942, due to the occupation of Norway by Germany. In the subsequent year, all prizes were awarded except those for literature and peace.

The Nobel Foundation is exempt from all taxes in Sweden (since 1946) and from investment taxes in the United States (since 1953).

Two laureates have voluntarily declined the Nobel Prize. In 1964, Jean-Paul Sartre was awarded the Literature Prize but refused, stating, "A writer must refuse to allow himself to be transformed into an institution, even if it takes place in the most honourable form.

In 1968, Sweden's central bank Sveriges Riksbank celebrated its 300th anniversary by donating a large sum of money to the Nobel Foundation to be used to set up a prize in honour of Alfred Nobel.

The following year, the Sveriges Riksbank Prize in Economic Sciences in Memory of Alfred Nobel was awarded for the first time. The Royal Swedish Academy of Sciences became responsible for selecting laureates. The first laureates for the Economics Prize were Jan Tinbergen and Ragnar Frisch,The Board of the Nobel Foundation decided that after this addition, it would allow no further new prizes.

Lê Đức Thọ, chosen for the 1973 Peace Prize for his role in the Paris Peace Accords, declined, stating that there was no actual peace in Vietnam.

Since the 1980s, the Foundation's investments have become more profitable and as of 31 December 2007, the assets controlled by the Nobel Foundation amounted to 3.628 billion Swedish kronor (c. US$560 million).

Youngest person ever to receive a Nobel Prize was Malala Yousafzai; at the age of 17, received Nobel Peace Prize (2014).

Oldest person ever to receive a Nobel Prize was John B. Goodenough; at the age of 97, received Nobel Prize in Chemistry (2019).