Anna Jagiellon was born on 18 October 1523 to the Polish king and queen, Sigismund I the Old and Bona Sforza. She spent most of her childhood in Kraków. Twice, from June 1533 to November 1536 and from April 1540 to June 1542, Anna and two of her sisters were left alone in Kraków while the rest of the family was in Lithuania. That meant that the three sisters grew closer (they separated only in 1556 when Sophia Jagiellon married Henry V, Duke of Brunswick-Lüneburg), but grew more distant from their elder brother Sigismund II Augustus. Her early life was rather mundane. She embroidered and sewed, played chess and dice, was involved in works of charity and fulfilled her obligations as a princess. She also received an education – she was fluent in Italian and knew Latin – but her teachers are unknown.
The issue of marriage of the three Jagiellon sisters was neglected by both their father and their mother. Only after the death of Sigismund I the Old in 1548, the first serious candidate to husband emerged – Albert Alcibiades, Margrave of Brandenburg-Kulmbach, but he was a Hohenzollern and a Protestant, had debts and a temper. In summer 1548, after a conflict with Sigismund II Augustus over his secret marriage with Barbara Radziwiłł, Queen mother Bona and her three unmarried daughters moved to Mazovia, mainly Warsaw and Ujazdów Castle. Sigismund II Augustus did not attend to the issue of his sisters' marriages. In 1550, Bona attempted to negotiate marriage with Charles Victor or Philip, sons of Henry V, Duke of Brunswick-Lüneburg, or Ernest of Bavaria. After a family meeting in May 1552 in Płock, Sigismund considered marrying his sisters to King Gustav I of Sweden, John Frederick II and Johann Wilhelm of Saxony, and John Albert I, Duke of Mecklenburg, but he lacked resolve and consistency.
Finally in January 1556, Queen Bona managed to arrange a marriage for Sophia Jagiellon. A month later, Bona Sforza departed to her native Italy leaving her two unmarried daughters alone in Warsaw. After about a year, Sigismund II Augustus brought his sisters to Vilnius where they became close to his third wife Catherine of Austria. Even though Anna was already in her mid-thirties, Sigismund investigated marriage proposals. Widowed Ferdinand I, Holy Roman Emperor, did not want to remarry; his unmarried son Charles II (born 1540) was too young; Tsar Ivan the Terrible was deemed not beneficial for Poland–Lithuania; John Frederick, Duke of Pomerania, did not want an alliance with Poland as it would have drawn the Duchy of Pomerania into the Livonian War.
King Eric XIV of Sweden was personally more interested in pursuing marriage with Queen Elizabeth I of England, but he sought an alliance with Poland and suggested his step-brother John, Duke of Finland. John agreed, but asked for Anna's younger sister Catherine. It was against custom for a younger sister to marry first, therefore their wedding was postponed. Three more grooms were proposed for Anna: Danish prince Magnus was supposed to become a bishop and, therefore, celibate; the last Master of the Livonian Order Gotthard Kettler was not of royal blood and his control of Livonia was tenuous; John proposed his younger brother Magnus, Duke of Östergötland, and Sigismund II Augustus agreed to the double Polish–Swedish alliance. However, at the last minute, only John arrived to the wedding in Vilnius. The court demanded that John married Anna, but John insisted on Catherine. Sigismund II Augustus, who needed Swedish troops and money in the Livonian War, relented if Anna did not protest. Though it must have been humiliating for Anna, she agreed and Catherine married John on 4 October 1562.
Anna moved to the Royal Castle in Warsaw as Vilnius was not safe due to the Livonian War and lived there for about ten years. There she lived alone with a court of about 70 people; she spent her time praying, embroidering or sewing (many of her works survived to this day), and corresponding with her sisters. Her brother visited her annually when he attended sessions of the general sejm in Warsaw. Even though Anna was already in her forties, marriage proposals continued to come in. In 1564, Reichard, Count Palatine of Simmern-Sponheim, proposed but perhaps was deterred by her relatively small dowry of 32,000 Polish red złoty. In 1568, Sophia Jagiellon proposed Eberhard, eldest son of Christoph, Duke of Württemberg, but he died the same year. In 1569, a project emerged to marry Anna to Barnim X, Duke of Pomerania. Pomerania demanded that Anna would bring eight border territories as her dowry, which was unacceptable to Poland. In 1572, Sophia proposed Albert Frederick, Duke of Prussia, but Sigismund II Augustus refused.
In July 1572, her brother Sigismund II Augustus died, leaving the throne to the Polish–Lithuanian Commonwealth vacant. His death changed her status from a socially neglected spinster to heiress of the Jagiellon dynasty. Sigismund left all the wealth of the Jagiellon dynasty to his three sisters. However, Polish nobles did not allow a private person to inherit royal possessions. Thus, Anna received only a small portion of her inheritance; it still made a very rich woman. In general, her life became dictated by Polish nobles, who sought to keep her out of the political arena. She was ordered to leave Warsaw and traveled to Piaseczna, Płock, Łomża.
Jean de Monluc, Bishop of Valence, offered the French prince Henry de Valois to the electors of the Commonwealth as the next king. Among other things, Montluc promised the electors that Henry would marry Anna to maintain the dynastic tradition. It seems that, despite nobility's efforts to keep her out of politics, Anna learned of Henry's offer in spring 1573 and became his strong supporter. She was flattered that Henry "cared for her and not for the Kingdom". With her support, Henry was elected as King of Poland on 11 May 1573 and officially crowned on 21 February 1574. Due to an oversight (whether intentional or unintentional), Henrician Articles (Henry's pre-election treaty) did not include the promise to marry Anna and so he delayed. When it became apparent that Henry would not marry her, Anna was humiliated. In June 1574, Henry left Poland to assume his new duties as King of France and by May 1575 the Parliament of the Commonwealth had removed him as their monarch.
During the second interregnum, Anna assumed the unprecedented but politically important title of infanta, mirroring the Spanish custom and highlighting her dynastic status. Poland did not recognize the status of crown prince since, technically, the monarchy was not hereditary but elective. She referred to herself as Anna, by the Grace of God, Infanta of the Kingdom of Poland (Latin: Anna Dei Gratia Infans Regni Poloniae). She wanted to marry and become Queen of Poland, but deceived by the French, she was a lot more careful and did not voice her support publicly. She was skeptical of marriage proposals by Archduke Ernest of Austria, Alfonso II d'Este, Duke of Ferrara, or Frederick IV of Liegnitz. In December 1575, Jan Zamoyski proposed to elect Anna. However, a woman could not rule and needed to marry and so Stephen Báthory, Voivode of Transylvania, was proposed as her husband. On 15 December 1575, in Wola near Warsaw, Anna and Stephen Báthory were elected as co-rulers of the Polish–Lithuanian Commonwealth. Lithuanian delegation did not participate in the election and did not recognize its results. Only on 29 June 1576, already after the coronation ceremony, Lithuanian nobility agreed to recognize Anna and Báthory.
Anna supported Báthory with money for weapons, but refused to get married per procura. On 28 February 1576, Anna entered Kraków as the officially elected Queen; Báthory joined her on 23 April. On 1 May they were married and crowned at Wawel Cathedral. Anna was forced to surrender inheritance from her brother; in return she received some of his properties for her lifetime, Mazovian properties that once belonged to her mother, treasury kept at Tykocin, and a one-time payment of 60,000 gold coins. She also received income from Wieliczka Salt Mine and interest on her mother's loan to Philip II of Spain (the loan was never fully repaid and is known as Neapolitan sums). She was supposed to share that interest with her sister Catherine Jagiellon, but apparently never did.
The marriage was consummated and there were rumors that Anna had not had her menopause and thus could still conceive, but it was a formal affair. The couple was distant and would see each other only a few weeks a year when Báthory, generally preoccupied with the Livonian War, attended general sejm in Warsaw. It visibly upset Anna who sought a closer personal relationship and greater political influence. It was also Báthory's loss who failed to gain a valuable political ally. Anna spent most of her time in Warsaw and Ujazdów Castle. There were rumors that Báthory might seek a divorce so he could marry a younger woman and father an heir; there were also rumors that he had an illegitimate son. This further humiliated and alienated Anna who even approached anti-Báthory groups and opposed his Livonian campaign.
Removed from politics and receiving substantial income, Anna sponsored several large constructions. She completed reconstruction of the Royal Castle in Warsaw and the Sigismund Augustus Bridge. She built the Bridge Gate to protect the newly built wooden bridge from fire. She also completed the Ujazdów Castle that was started by her mother; it had a large park and a menagerie. In the Wawel Cathedral, she funded the tomb monument of her brother King Sigismund II Augustus in the Sigismund's Chapel (1574–75 by Santi Gucci). The chapel received a new gilded roof in 1584. After her husband's death, she refused to allow his burial in the Sigismund's Chapel; perhaps it was her retribution for the distant marriage as traditions dictated that husband and wife should not be separated in death. Stephen Báthory was buried in the Chapel of the Blessed Virgin Mary. However, Anna did order his tomb monument in 1589. Around the same time she built her own tomb in the Sigismund's Chapel. She also built the tomb of her mother Bona Sforza in the Basilica di San Nicola in Bari (1589–95).
With the death of her husband in December 1586, Anna had an opportunity to claim the political power in the Commonwealth for herself as she was elected as Queen in her own right. If needed, a new husband could be found. However, she did not even attempt it. Instead, she resolved to promote her niece Anna Vasa or her nephew Sigismund Vasa, the only children of her beloved sister Catherine Jagiellon and King John III of Sweden. Her initial plan, formulated while her husband was still alive, was to wed Anna Vasa and one of the nephews of Stephen Báthory and promote the couple to the throne. However, this plan did not gain support among the nobility. She then planned to sponsor Sigismund Vasa to the throne. Initially, King John III did not want to let his sole heir out of his sight, but Anna managed to convince him. As a back up plan, she pursued marriage between Anna Vasa and Maximilian III, Archduke of Austria, the other likely candidate to the throne. In her campaigns, she wrote numerous letters and used her wealth. She also gained crucial support from Jan Zamoyski, who was married to Báthory's niece Griselda Báthory and held his own ambitions for the throne. Perhaps both Anna and Jan thought that they could easily control the inexperienced 20-year-old.
Sigismund Vasa was elected as King on 19 August 1587. He and his sister Anna Vasa arrived to Poland in October 1587. After the coronation and the brief War of the Polish Succession, Anna and her niece settled in Warsaw while Sigismund spent most of his time in Kraków. It became clear that he would not allow his aunt to interfere in politics. Despite her disappointment, Anna became attached to her nephew. She participated in his wedding with Anne of Austria and baptism of their firstborn Anna Maria. After the death of John III of Sweden in November 1592, Sigismund Vasa spent about a year in Sweden. During that time, the newborn Anna Maria was entrusted to Anna's care. In July 1595, she was the godmother of Władysław Vasa, the future King of Poland.
Anna died in Warsaw on 9 September 1596 as the last Jagiellon.