The highway runs through (West to East): Riedstadt, Griesheim, Darmstadt, Dieburg, Münster, Hesse, Babenhausen, Großostheim, Aschaffenburg, Goldbach, Bavaria, Hösbach, Laufach, Rechtenbach, Lohr, Gemünden am Main, Karlstadt am Main, Arnstein, Werneck, Bergrheinfeld, Schweinfurt, Schonungen, Theres, Haßfurt, Zeil am Main, Ebelsbach, Eltmann, Viereth-Trunstadt, Bischberg and Hallstadt.

In Darmstadt the B26 meets the North-South Highway of Bundesstraße 3, B3.

As a replacement for the crossing of the Darmstadt city center, a north eastern bypass is planned.

It should relieve the downtown area from car and truck traffic.

The planned route was to connect the Darmstadt Ostbahnhof and the Martin-Luther-King-ring.