The payload of the Faint Intergalactic Redshifted Emission Balloon (FIREBall-2), the second generation of the FIREBall instrument (PI: C. Martin, Caltech), has been calibrated and launched from the NASA Columbia Scientific Balloon Facility in Fort Sumner, New Mexico. FIREBall-2 was launched for the first time on the September 22, 2018, and the payload performed the very first multi-object acquisition from space using a multi-object spectrograph. Our performance-oriented paper presents the calibration and last ground adjustments of FIREBall-2, the in-flight performance assessed based on the flight data, and the predicted instrument’s ultimate sensitivity. This analysis predicts that future flights of FIREBall-2 should be able to detect the HI Lyα resonance line in galaxies at z  ∼  0.67, but will find it challenging to spatially resolve the circumgalactic medium.