Analysis of Ricin Subunits by High-Resolution Acidic Native Gel Electrophoresis

Order of Publishing in Issue: 
4
Volume :5
Issue :4
October, 2011
Page No: 
1410-1413
Authors: 
Payal Puri, Om Kumar*, Krishna Chaturvedi and Ramesh Kaul
Address: 
Division of Pharmacology and Toxicology, Defence Research and Development Establishment Jhansi Road, Gwalior-474002, India

Abstract
Ricin is a potent toxic castor bean glycoprotein comprising of two subunits, A chain and B chain. We studied ricin subunits in an acidic electrophoresis system followed by electro blotting. To the best of our knowledge, there is no study reported for characterization of ricin subunits using acidic native PAGE. Sample preparation steps for acidic native PAGE were optimized and adequate electrophoretic conditions were developed for achieving a complete separation of ricin subunits. Ricin subunits were separated and characterized at acidic pH (2.9 and 4.3) and their immunological detection was made using polyclonal antibodies. Our findings showed that this method results in sharper band and therefore high resolution than SDS-PAGE and also the following protein transfer is highly efficient and rapid.

Keywords: 
Ricin, Acidic non-denaturing PAGE, Electro blotting, Resolution.
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