The flag was flown from the large public flagpole in San Francisco's Castro District (where the rainbow flag usually flies) for the first time on 19 and 20 November 2012 in commemoration of the Transgender Day of Remembrance.

Transport for London also flew the flag from London Underground's 55 Broadway Headquarters for the 2016 Transgender Awareness Week. In the United Kingdom, Brighton and Hove council flies this flag on the Transgender Day of Remembrance. The stripe in the middle is white, for those who are transitioning or consider themselves having a neutral or undefined gender. The stripes next to them are pink, the traditional color for baby girls. The stripes at the top and bottom are light blue, the traditional color for baby boys. Helms describes the meaning of the transgender pride flag as follows: The flag's design represents the transgender community, and consists of five horizontal stripes: two light blue, two pink, and one white in the center.

The flag was created by American trans woman Monica Helms in 1999, and was first shown at a pride parade in Phoenix, Arizona in 2000. The most prominent transgender flag design is the 'Transgender Pride Flag', used as a symbol of transgender pride and diversity, and transgender rights.