Compositions and Methods for Improving Resolution of Biomolecules Separated on Polyacrylamide Gels
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Native Protein / Protein Complex Gel Electrophoresis
[0110] Gel and Cassettes: 3-12% polyacrylamide gradient gels were poured comprising acrylamide / bisacrylamide in a ratio of 25.5:1 in BisTris-Cl, pH 7.0 and 0.3 mM CHAPS detergent. The stacker was 3% acrylamide / bisacrylamide in a ratio of 25.5:1 and 0.05% w / v linear acrylamide (600,000 to 1,000,000 Daltons). A first gel was poured in a cassette in which the measurement of the gap in the middle of the cassette along the upper edge was 0.923 mm, and a second gel was poured in a cassette having a gap of 0.991 mm in the middle of the cassette along the upper edge. The interplate gaps at the left and right edges of these cassettes were between 1.01 mm and 1.02 mm, therefore the first gel cassette has a narrower middle region compared with the second gel cassette.
[0111] Sample Preparation: Bovine mitochondria were isolated as described previously (Rice, J. E. & Lindsay, J. G. (2002) “Subcellular fractionation of mitochondria” in Subcellul...
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