On-sky tests conducted with an astro-comb using the Potsdam Multi-Aperture Spectrograph (PMAS) at the 3.5 m Calar Alto Telescope are reported. The proposed astro-comb approach is based on cascaded four-wave mixing between two lasers propagating through dispersion optimized nonlinear fibers. This approach allows for a line spacing that can be continuously tuned over a broad range (from tens of GHz to beyond 1 THz) making it suitable for calibration of low- medium- and high-resolution spectrographs. The astro-comb provides 300 calibration lines and his line-spacing is tracked with a wavemeter having 0.3 pm absolute accuracy. First, we assess the accuracy of Neon calibration by measuring the astro-comb lines with (Neon calibrated) PMAS. The results are compared with expected line positions from wavemeter measurement showing an offset of 5–20 pm (4%–16% of one resolution element). This might be the footprint of the accuracy limits from actual Neon calibration. Then, the astro-comb performance as a calibrator is assessed through measurements of the Ca triplet from stellar objects HD3765 and HD219538 as well as with the sky line spectrum, showing the advantage of the proposed astro-comb for wavelength calibration at any resolution.