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A Docile Method of Isolation and Purification of Riboflavin Binding Protein (Rfbp) from Peahen (Pavo cristatus) and Domestic Fowl (Gallus gallus)
Riboflavin binding protein (Rfbp) was isolated from domestic fowl (Gallus gallus) and peahen (Pavo cristatus) egg-white and egg-yolk. The protein was purified in two steps, DEAE-Sephadex A-50 ion exchange chromatography and eluted with phosphate buffer pH 7.3 containing 0.5 M sodium chloride. The final purification of protein was achieved on Sephadex G-100. The purity of the protein was judged on cylindrical and slab gel electrophoresis, SDS-PAGE technique. Sephadex G-100 fraction Rfbp moved as a single band both on the Slab and Cylindrical gels. Comparison of the mobility of Rfbp with that of the standard molecular weight marker proteins revealed with that the Rfbp had a molecular weight close to 29,000 kd. Interestingly, hen egg-white Rfbp and peahen egg- white, yolk Rfbp had the same molecular weight as revealed by the SDS-PAGE. This is a novel approach for the study of riboflavin binding protein purified from different avian eggs in two steps and studied electrophoretic characterization with standard molecular weight marker.
Rfbp Purification Method, Peahen, Hen, Egg White-yolk, SDS, PAGE/native
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