Recovery of native proteins from preparative electrophoresis gel slices by reverse polarity elution.
A technique for high yield recovery of native, biologically active proteins from preparative polyacrylamide gel slices by reverse polarity elution is described. No apparatus other than the standard slab gel electrophoresis system is required. Several proteins have been recovered in biologically active form at a 90% yield, in quantities ranging from 0.4 mg to 4.2 mg. The method is effective with both small (9,000 dalton) and large (186,000 dalton) polypeptides. Both simple and complex proteins are recovered intact. For example, the copper-zinc and manganese superoxide dismutases from crude soybean extracts are active upon recovery. Similarly, the vitamin D-dependent calcium binding proteins from rat kidney and intestine are isolated by this method in homogeneous, active form.
D000818:Animals; D001234:Aspergillus niger / Q000201:enzymology; D003574:Cytochrome c Group / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005949:Glucose Oxidase / Q000302:isolation & purification; D006736:Horses; D007422:Intestines / Q000032:analysis; D007668:Kidney / Q000032:analysis; D011506:Proteins / Q000302:isolation & purification*; D051381:Rats; D064030:S100 Calcium Binding Protein G / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D013025:Glycine max; D013482:Superoxide Dismutase / Q000302:isolation & purification
D003574:Cytochrome c Group; D011506:Proteins; D064030:S100 Calcium Binding Protein G; D012710:Serum Albumin, Bovine; D005949:Glucose Oxidase; D013482:Superoxide Dismutase