The founding date of the Nikolo-Volosovo Monastery remains unknown. The first mentions of the Volosovo monastery date back to the 15th century. In Russia, there are many places where ancient shrines of the pagan god Veles were replaced by Nikolai churches and monasteries. One such place is located near Vladimir, in the village of Volosovo. The nuns of the monastery pass down a tradition that the monastery was originally built on the site of a destroyed shrine of Veles. According to legend, the Nikolai church was first erected on a hill, at the site of the pagan shrine, but the miraculous image of Nicholas of Myra repeatedly disappeared from the church and was found in the lowland by the Kolotchka River, hanging from a tree by its hair. Eventually, the church was moved to the place chosen by the icon, and soon a monastery arose there, and the village was named Velesovo.

In the 17th century, the first stone structure, the Sergiev Church, was built in the monastery. In addition to the main altar in honor of St. Sergius of Radonezh, there was a side chapel in honor of the Equal-to-the-Apostles Constantine and Helena. In the early 18th century, Peter I began to reduce the number of monasteries, with the income intended for state needs. Small monasteries with fewer than 30 monks were merged with others or closed. In 1722, by decree of the Holy Synod, the Lyubetsky Assumption and Protection Monasteries on the Nerl River were assigned to the Nikolai Volosovo Monastery. From 1725 to 1727, the Kosmin Monastery was attached to the Volosovo Monastery. Under the abbess Paul, the cathedral Nikolai church was built.

In 1763, the Protection gate church was erected, which remained unblessed for a long time and gradually fell into disrepair. The walls of the church cracked due to the poor quality of the material. In the 1890s, the church was restored thanks to peasant Yakov Ivanovich Busurin, who helped the monastery. On September 21, 1893, the church was solemnly consecrated by Bishop Tikhon of Murom. From the mid-19th century until 1909, the monastery was inactive. On December 18, 1843, by decree of the Holy Synod, the Nikolo-Volosovsky Monastery was abolished due to the dilapidation of the buildings. The brothers were transferred to the Holy Bogolyubov Monastery, but the Volosovo monastery did not remain empty: there were an hieromonk, two novices, and a servant who took turns coming from Bogolyubovo to conduct services. The idea of reviving the monastery arose in the 19th century among church ministers and laypeople. Saint Theophan, who was on the Vladimir see, proposed to the Synod in 1865 to restore the monastery as a missionary center. Although his project was not realized, it foreshadowed the revival of the monastery. In 1909, the Volosovo Men's Monastery became a women's monastery. On February 18, 1911, nun Thais was appointed abbess.

In 1927-1928, Sergius, transferred from Vladimir, served in the Nikolai church. He was arrested three times and shot in 1937. Former residents lived near the monastery in houses where local residents took them in. From the 1930s to the 1980s, the monastery buildings were used by the collective farm, with a store and bakery located in the Nikolai church, while other buildings served as warehouses and housing for collective farm workers.

The monastery was revived in 1992 by the sisters of the Bogolyubsky Women's Monastery. In 1993, the semi-ruined buildings of the Nikolo-Volosovsky Monastery became a dependency of the Suzdal Protection Women's Monastery, and since 1996, it has been an independent monastery.