Purification of Maltose-Binding Protein from E. coli Periplasm

 

Judith Levin

Tested Studies in Laboratory Teaching, 1997, Volume 18

Views: 41 Downloads: 68^a

Abstract

By correlating the protein concentration of various fractions from E. coli with the appearance of samples on a gel, this exercise introduces beginning students to the quantitative “bookkeeping” involved in protein purification. A periplasmic fraction is isolated by osmotic shock, and affinity chromatography with amylose resin is used to purify maltose-binding protein. Visualization of fractions by SDS-polyacrylamide gel electrophoresis allows students to evaluate the qualitative success of the purification and to estimate the subunit molecular weight of the purified protein.

Keywords:  cell fractionation, protein purification, affinity chromotography, protein determination, standard curve, SDS-PAGE, osmotic stress

Boston University (1996)

^aAs of September 1, 2026