[3dem] adsorption problems?

Reinhard Rachel Reinhard.Rachel at biologie.uni-regensburg.de
Sun Oct 13 23:34:10 PDT 2013

Hi Michael,
this might be a problem of "adsorption" to (Carbon coated? hydrophilized?) surfaces / grids. 
But, more likely, you have a biological / biochemical 'phenomenon' - is it possible that this "single subunit" acts to stabilize the whole complex? Have you tried other means for separation (not only gradient centrifugation, but: native gel electrophoresis, or any chromatography)? And, you may combine gradient centrifugation with a mild fixation step (GRAFIX; Kastner et al, in Nature Methods - 5, 53 - 55 (2008) Published online: 23 December 2007; | doi:10.1038/nmeth1139 ) 
You even may try to do a quick chemical fixation (0.1 to 1% GA; 5 min) before you apply your sample onto the grid. And, the stability of the whole complex +/- subunit may depend and vary on the pH: do you only use UAc, or did you try another negative staining solution (PTA or whatsoever)? 
lots of things to consider. 
kind regards,

Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel at biologie.uni-regensburg.de
office: VKL 3.1.29 
Last microscopy conference: 
MC2013 in Regensburg, Germany
Next microscopy conference:
18th IMC 2014 in Prague, Czech Rep.

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