Below is a stack (977 MB) of 31,592 images (MRC/CCP4 format) of N-ethyl maleimide sensitive factor (NSF ). The images were recorded on a Philips CM12 or FEI Tecnai F20 FEG electron microscope under low-dose conditions and between 2.0 and 4.0 µm underfocus. The pixel size of the final reconstruction is 3.5 Å. Alignment parameters determined by Frealign for each particle in the stack can also be found below, as well as a reconstruction of NSF (also MRC/CCP4 format; a B-factor of -1900 Å2 and an 11 Å low-pass filter were applied using the program bfactor to sharpen the density) and a script to calculate a reconstruction using Frealign.
Please note that the handedness of the reconstruction calculated using the attached parameter files will be WRONG. The reconstruction must be mirrored (for example, with the MRC program label) to have the correct handedness. The reconstruction attached below has already been corrected.