The gradient fixation method, in which a glycerol gradient centrifugation is performed in the presence of a cross-linker, helps to identify interactions between proteins that bind transiently to multi-subunit complexes. The use of fixation reagent
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Here, we describe the utilization of Grafix (Gradient Fixation), a glycerol gradient centrifugation in the presence of a crosslinker, to identify interactions between splicing factors that bind transiently to the spliceosome complex.