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The Chemical Educator
ISSN: 1430-4171 (electronic version)
Abstract Volume 17
(2012) pp 071-072
DOI 10.1007/s00897122415a
Ion Exchange Chromatography of Proteins Using Microspin Columns: An Undergraduate Biochemistry Laboratory
Eathan McIntyre, Tammy Melton, and Paul C. Kline*
Department of Chemistry, Middle Tennessee State University,
Murfreesboro, TN 37132,
paul.kline@mtsu.edu
Received September 21, 2011. Accepted March 27,
2012.
Published: 25 May 2012
Abstract. Details and instructions for an undergraduate biochemistry laboratory are described. Students carry out a separation of ˛-glucosidase and cytochrome c on microspin ion exchange columns. Students are challenged to predict the separation of the proteins based on their differences in isoelectric point, and the cationic nature of the columns. The concentration of the proteins is determined by absorption spectroscopy at 280 nm, and also via a protein dye microassay. Students prepare and explain an elution profile of the fractions obtained from the columns. A description of instructions to students is included.
Key Words: Laboratories and Demonstrations; biochemistry chemistry
(*) Corresponding author. (E-mail: paul.kline@mtsu.edu)
Article in PDF format (87 KB) HTML format
Supporting Materials:
The following information is available as supporting material: (i) Student handout; (ii) Typical student data (77 KB).
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