(a) Brij-lysed erythrocyte suspension (lysate, = L) layered onto a 25-40-50 % glycerol step gradient in a 200ml conical plastic centrifuge bottle. (Note - lower part of tube actually contains clear gradient solutions; it appears a bit cloudy due to reflection of substrate). (b) Same after centrifugation; hemoglobin (Hb) and other soluble components remain in upper part of gradient, while cytoskeletons have accumulated in the 50% glycerol layer at the bottom.