The gallery's history starts in 1856 when the Muscovite merchant Pavel Mikhailovich Tretyakov acquired works by Russian artists of his day with the aim of creating a collection, which might later grow into a museum of national art.

In 1892, Tretyakov presented his already famous collection of approximately 2,000 works (1,362 paintings, 526 drawings, and 9 sculptures) to the Russian nation.

The collection contains more than 130,000 exhibits, ranging from the Theotokos of Vladimir to the monumental Composition VII by Wassily Kandinsky and the Black Square by Kazimir Malevich.

[5] In May 2023, the Tretyakov Gallery refused to hand over one of its most famous icons, Andrei Rublev's Trinity, to the Russian Orthodox Church.

[6] In June 2023 the icon was transferred to Moscow's main cathedral despite the museum's protests on the personal order of Russian President Vladimir Putin.

The official opening of the museum called the Moscow City Gallery of Pavel and Sergei Tretyakov took place on 15 August 1893.

With Grabar's active participation in the same year, the State Museum Fund was created, which up until 1927 remained one of the most important sources of replenishment of the gallery's collection.

Later, the church was connected to the exposition halls and a top floor was built which was specially designed for exhibiting a painting by Alexander Andreyevich Ivanov,The Appearance of Christ Before the People (1837–1857).

Paintings were rolled on wooden shafts, covered with tissue paper, placed in boxes, and sheathed with waterproof material.

From 1986 to 1995, the Tretyakov Gallery in Lavrushinsky Lane was closed to visitors to accommodate a major renovation project to the building.

The grounds of this branch of the museum contain a collection of Socialist Realism sculpture, including such highlights as Yevgeny Vuchetich's iconic statue Iron Felix (which was removed from Lubyanka Square in 1991), the Swords Into Plowshares sculpture representing a nude worker forging a plough out of a sword, and the Young Russia monument.

Near the gallery of modern art there is a sculpture garden called "the graveyard of fallen monuments" that displays statues of former Soviet Union that were relocated.