This study investigated the effectiveness of electrical muscle stimulation (EMS) with resistance exercise training (ERT) and resistance exercise training (RT) on physical and brain function in middle-aged and older women. Method: Forty-eight participants were randomly allocated into three groups: (i) ERT (n = 16), (ii) RT (n = 16), and (iii) control group (n = 16). The intervention session was 50 min long and performed three times/week for four weeks. The ERT group performed quadriceps setting, straight leg raises, and ankle pump exercises while constantly receiving EMS on their quadriceps muscle on both legs. The RT group performed the same exercise without EMS. Physical function was measured using skeletal muscle mass index (SMI), handgrip strength, gait speed, five times sit-to-stand test (FTSS) and timed up-and-go test (TUG). Brain function was assessed with electroencephalogram measurement of whole brain activity. Results: After four-week intervention, significant improvements were observed in SMI (p < 0.01), phase angle (p < 0.05), and gait speed (p < 0.05) in the ERT group compared to the control group. ERT also increased muscle strength (p < 0.05) and mobility in lower limbs as observed in FTSS and TUG tests (p < 0.05) at post-intervention compared to the baseline. In the ERT group, significant positive changes were observed in Beta1 band power, Theta band power, and Alpha1 band whole brain connectivity (p < 0.005) compared to the control group. Conclusions: Our findings showed that ERT can improve muscle and brain function in middle-aged and older adults during a four-week intervention program whereas significant improvements were not observed with RT. Therefore might be one of the feasible alternative intervention to RT for the prevention of muscle loss whilst improving brain function for middle-aged and older population.