The East Bohemian Gallery did not have an opportunity to develop any previous historical collections. Since the early 1950s, it has based its collections on grants from the National Gallery and the Ministry of Education and Culture, on donations and of course its own acquisitions. As it is the case in other regional and state galleries, these acquisitions have been determined not only by the amount of financial resources, but also mainly by the personality of the director and collection curator.
The present-day gallery collections comprise of a collection of Czech art of the 19th century, with paintings by A._Machek, A._Piepenhagen, J._Mánes, A._Kosárek, A._Bubák, A._Chittussi, A._Waldhauser, J._Mařák, etc., and above all of a collection of Czech art of the 20th century and a collection of regional art. The collections include a large set of landscape paintings from the end of the 19th century up to the contemporary tendencies. In this set, a special attention is paid to the works by landscape painters from the Iron Mountains and the Highlands. The most noteworthy sets also present works of the civilism and social art of the 1920s, works by members of the Skupina (Group) 42, from the 1968 and 1969 Artchemo Symposia and others. As for the authors' collections, there are sets of works by E._A._Pitterman-Longen, J._Honsa, A._Hudeček, J._Panuška, F._Kaván, S._Máchal, T._F._Šimon, J._Bauch, F._Muzika, J._Smetana, M._Holý, J._Kotík, K._Lhoták, J._Konůpek, E._Filla, F._Tichý, B._Reynek, J._Grus, O._Janeček, J._Trampota. Another excellent set is a collection documenting Czech art of the 1960s - M._Medek, S._Podhrázský, V._Preclík, J._Šerých, M._Grygar, J._Hilmar, O._Slavík, E._Kmentová, J._Kolář, V._Jarcovják, F._Ronovský, S._Podhrázský, Z._Fibichová, K._Malich, V._Janoušková, F._Kyncl, D._Chatrný, J._Hendrych, J._Pešicová, F._Štorek and others.
The painters from the Pardubice region are well represented in the collections as well. The gallery also manages the largest set of works by Jaroslav Grus, a native of Pardubice. The set consists of 169 paintings, 82 drawings and 22 graphic sheets. This collection was created during the years 1974 - 1978. It was intended for the author's permanent exhibition open in the Dům U Jonáše until 1991. In 2002, the purchase of 83 works from the former collection of Czech Telecom enriched the collection of regional artists with works by F._V. Danihelka, A._Hajn, J._Hegr, M._Horáková, W._Horný, L._Jandová, J._Klápště, V._Komárek, B._Kopecký, J._Kopecký, J._Kříž, F._Kyncl, J._Lacina, S._Malý, J._Mědílek, J._Nekovář, L._Novák, B._Novotný, L._Pejchl, J._Procházka, K._J._Sigmund, Z._Sigmund, J._Ščerbakov, M._Ščigol, V._Truhlářová, V._Vokolek and L._Zívr.
In total, the East Bohemian Gallery in Pardubice has 14.961 collection objects including negatives and photographs. The collection of graphic art is ample; it amounts to 4.072 graphic sheets. The collection of paintings has 2018 works and the collection of drawings 1484 works. We can find 467 exhibits in the collection of sculpture. In 1994, the gallery started to purchase an extensive set of negatives and photographs by Jiří_Toman (1924 - 1972). The set documents the works of an outstanding personality from the Pardubice culture of the 1960s. At present, the gallery keeps 6.801 negatives and 119 photographs by this author.