Trembling aspen (Populus tremuloides) is one of the major species within Populus, a predominant genus of hardwoods in North America. However, its utilization has been limited to pulp and paper or wood-based composite boards. This study aimed at evaluating the major physical and mechanical properties of trembling aspen lumber, with an ultimate objective of using this species to produce engineered wood products (EWPs). The testing materials consisted of 2 × 4 (38 mm × 89 mm) trembling aspen lumber pieces in lengths of 8, 10, and 12 feet (2.44, 3.05, and 3.66 m) with two visual grades, select structural (SS) and No. 2. Machine Stress-Rated (MSR), and longitudinal stress wave (LSW), edgewise third-point bending (EWB), and axial tension tests were conducted on the lumber. It was found that, (1) by increasing the maximum knot size by a half-inch from one-quarter inch, the minimum modulus of elasticity (MOE) measured using the MSR, the mean, and the fifth-percentile ultimate tensile strength (UTS) decreased by about 8.8%, 20.1%, and 29.8%, respectively. (2) Approximately 44% of the trembling aspen lumber met the 1450f-1.3E grade for MSR lumber, and 62% qualified for the 1200f-1.2E grade. (3) There was a great potential for manufacturing cross-laminated timber (CLT) of grade E3, with a rejection rate of about 29%. (4) The mean UTS and MOE values of the SS-grade trembling aspen lumber were 22.88 MPa and 9519 MPa, respectively, being 25.5% and 11.3% lower than that of Spruce–Pine–Fir (S-P-F) lumber. The fifth-percentile UTS and MOE values were 11.57 MPa and 7404 MPa, respectively, marking a decrease of 13.3% and 1.5% compared to the S-P-F lumber. (5) The oven-dried specific gravity (SG) of the trembling aspen wood was 0.40, which was about 3.5% larger than the value provided in the Wood Handbook.