The first ordained rabbi was Rabbi Michael Fried, who came to the Synagogue in 1898, directly from the Jewish Theological Seminary. Under his tenure, the Synagogue moved to its Craft Avenue site in 1906. He was succeeded by Rabbi Rudolph Coffee, who served the Congregation until 1915. After Rabbi Coffee left the Tree of Life for the West Coast in 1915, he was succeeded by Rabbi Morris M. Mazure, who remained with the Congregation until 1921.

Rabbi Herman Hailperin came to the Tree of Life in 1922, and served the Congregation for a period of fifty years. Rabbi Hailperin became Rabbi Emeritus upon the arrival of Rabbi Solomon M. Kaplan in 1968.

Rabbi Alvin K. Berkun began his tenure at Tree of Life in 1983. Rabbi Avi Friedman joined Rabbi Berkun in 1999 and served as Rabbi until the summer of 2005.

Rabbi Stephen Listfield currently serves as the eighth Rabbi at Tree of Life Congregation.