Supplementary Figure 3: General procedure for establishing Intensity vs. Charge regression model shown for AaLS-neg, (z=149). | Nature Methods

Supplementary Figure 3: General procedure for establishing Intensity vs. Charge regression model shown for AaLS-neg, (z=149).

From: Resolving heterogeneous macromolecular assemblies by Orbitrap-based single-particle charge detection mass spectrometry

Supplementary Figure 3

A Single particle centroids, preprocessed and filtered as described in Supplementary Fig. 8, were binned in m/z and smoothed. The resulting mass spectrum was used for conventional charge state assignment. B Centroids were filtered for charge state 149 by only allowing centroids within 80 Th of the top peak position to pass the filtering. C Filtered centroids were subjected to a kernel density estimation and the most abundant intensity as well as the FWHM of the distribution were extracted. D To obtain an equal weighted fit over all tested charges 200 random samples were drawn for each charge within two times the FWHM of the most abundant intensity. Assuming a normal distribution this contains more than 98 % of the centroids (FWHM = 2.35σ). E & F The same procedure was repeated for the other examined proteins and charges and used for establishing the linear regression model.

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