Purification of protein from a crude mixture through SDS-PAGE transfer method.
SDS-polyacrylamide gel electrophoresis (SDS-PAGE) transfer method was used for purification and enrichment of the protein from crude sample. Coomassie bluc/ZnSO4 stained protein band(s) containing intact polyacrylamide gel were loaded on to another polyacrylamide gel either alone or as pooled gel bands. Two/three bands were combined together and arranged tightly over one another, sealed with stacking gel and ran in another gel, which was quite useful for enrichment and purification of a particular protein from a complex mixture. Recovery of protein by gel transfer method was found to be 70% in case of ZnSO4 staining, whereas around 30% recovery was possible, following Coomassie blue staining. The method described here for purification of protein(s) from a complex mixture, following gel transfer procedure could be useful for further characterization of the desired protein.
Bhattacharyya
Dipankar
D
Department of Chemistry, Bose Institute, 93/1, A.P.C. Road, Kolkata 700 009, India.
Basu
Arindam
A
Sen
Parimal C
PC
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006041:Goats; D008297:Male; D012710:Serum Albumin, Bovine / Q000302:isolation & purification*; D013094:Spermatozoa / Q000737:chemistry*; D013194:Staining and Labeling; D013737:Testis / Q000737:chemistry
D012710:Serum Albumin, Bovine