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We evaluate the uncertainties of methane optimal estimation retrievals from single footprint thermal infrared observations from the Atmospheric Infrared Sounder (AIRS). These retrievals are primarily sensitive to atmospheric methane in the mid-troposphere through the lower stratosphere (~2 to ~17 km). We compare to in situ observations made from aircraft during the Hiaper Pole to Pole Observations (HIPPO), the NASA Atmospheric Tomography Mission (ATom) campaigns, and from the NOAA ESRL aircraft network, between the surface and 5-13 km, across a range of years, latitudes between 60 S to 80 N, and over land and ocean. After a global, pressure dependent bias correction, we find that the land and ocean have similar biases and that the reported observation error (combined measurement and interference errors) of ~27 ppb is consistent with the standard deviation between aircraft and individual AIRS observations. A single measurement has measurement (noise related) uncertainty of ~17 ppb, a ~20 ppb uncertainty from radiative interferences (e.g. from water, temperature, etc.), and ~ 30 ppb due to “smoothing error”, which is partially removed when making comparisons to in situ measurements or models in a way that account for this regularization. We estimate a 16 ppb validation error because the aircraft typically did not measure methane at altitudes where the AIRS measurements have some sensitivity, e.g. the stratosphere. Daily averaged AIRS measurements of at least 9 observations over spatio-temporal domains of < 1 degree and 1 hour have a standard deviation of ~17 ppb versus aircraft, likely because the observation errors from temperature and water vapor (for example) are only partly reduced through averaging. Seasonal averages can reduce this ~17 ppb uncertainty further to ~10 ppb, as determined through comparison with NOAA aircraft, likely because uncertainties related to radiative effects of temperature and water vapor can be reduced when averaged over a season.