Ankhesenamun was born into the royal family of the Eighteenth Dynasty. Her father Akhenaten promoted the Aten, the visible sun disk, and established his new capital at Akhetaten, now called Amarna. Her mother Nefertiti held a prominent place in royal ritual and art. Ankhesenpaaten appears among their daughters, alongside her older sisters Meritaten and Meketaten. The reliefs present the family participating in worship and in unusually intimate royal scenes. They reveal her place within the dynasty, although her exact birth date and the details of her childhood are not securely known.

The final years of Akhenaten's reign and the succession that followed remain difficult to reconstruct. The death of Meketaten is represented in the royal tomb at Amarna, but the cause of her death is uncertain; childbirth is one interpretation, not an established fact. The roles and sequence of the rulers Smenkhkare and Neferneferuaten are also debated. When Tutankhaten became king, Ankhesenpaaten was his wife. Their precise biological relationship depends on unresolved questions about his parentage, so it should not be presented as a completely settled family tree.

The couple's new names marked a political and religious change. Tutankhaten became Tutankhamun, while Ankhesenpaaten became Ankhesenamun, replacing the reference to Aten with one to Amun. During Tutankhamun's reign the court left Amarna for Memphis, and relations with the traditional religious centre at Thebes were restored. These changes placed the queen within the recovery of older institutions after her father's reforms. Her name was therefore more than a personal label: it also reflected the religious direction of the court she represented.

Objects from Tutankhamun's tomb show the royal couple together, including a box lid on which Ankhesenamun offers flowers to the king. Such images can feel strikingly personal, but they also belong to the symbolic language of Egyptian royal art and cannot reveal every detail of a marriage. Two small mummified children were also buried in the tomb and are generally regarded as Tutankhamun's daughters. Ankhesenamun is often proposed as their mother because she is his only securely known wife, but that identification is not independently certain. Their burials should not be described as physical remains of the queen herself.

After Tutankhamun died without a surviving heir, Ay succeeded him. Another episode, preserved in Hittite accounts of the reign of Suppiluliuma I, describes an Egyptian royal widow requesting one of the Hittite king's sons as her husband. The woman is called Dakhamunzu, a rendering of the Egyptian title meaning the king's wife rather than a personal name. Many scholars identify her with Ankhesenamun, while others propose a different queen. Suppiluliuma sent his son Zannanza, who died before the planned marriage took place. The Hittite account blames the Egyptians, but neither the exact circumstances nor a particular Egyptian perpetrator can be established with confidence.

A ring associated with the names of Ay and Ankhesenamun has been used to suggest that she later married him, but it does not establish the circumstances of such a marriage or prove coercion. Ay's wife Tey appears in his royal tomb. Ankhesenamun's later life and burial remain unresolved, and this absence of a clear record cannot by itself demonstrate murder. Horemheb and later rulers appropriated or dismantled monuments of the Amarna-era kings, but it is also misleading to say that every trace of the queen was destroyed. Her name survives on objects, including faience rings, and her images remain among the most memorable records of Tutankhamun's reign.