In Alabama, the Interstate Highway runs 80 miles (130 km) from I-65 in Montgomery northeast to the Georgia state line near Valley.

The Interstate also connects Montgomery with Tuskegee, Auburn, Opelika, and, indirectly, Phenix City and Columbus, Georgia.

The Interstate's final access to US 80 is via SR 186 (Wire Road), beyond which the freeway passes through a portion of the Tuskegee National Forest.

I-85 and US 29 continue northeast through the city to an interchange with Bent Creek Road, which provides access to Auburn University Regional Airport.

I-85 gains a second US Route concurrency at its four-ramp partial cloverleaf interchange with Gateway Drive, where US 280 joins from the north.

US 29 diverges from I-85 at the following interchange, and the Interstate leaves the city of Opelika beyond its junction with Andrews Road.

Southbound I-85 has a welcome center between CR 208 and the freeway's final interchange in Alabama, a junction with US 29 between the cities of Valley to the south and Lanett to the north.

[3] An extension of I-85 is proposed west from Montgomery to interchange with I-20 and I-59 just east of the Mississippi–Alabama state line,[5] where it will connect with I-20 and I-59 near Cuba.

[11] After years of stagnation, area leaders began calling for the construction of phase II of the project, which would extend the roadway to US 231/US 82 near Pike Road.

Pike Road Mayor Gordon Stone said the project is ready to move forward, but it cannot do so until it is approved for funding.