Ion exchange chromatography: Difference between revisions
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Ion-exchange [[Chromatography|chromatography]] separates [[Proteins|proteins]] according to their net charge. The stationary phase of the column will have a specific charge. | Ion-exchange [[Chromatography|chromatography]] separates [[Proteins|proteins]] according to their net charge. The stationary phase of the column will have a specific charge. | ||
For example negatively charged [[Proteins|proteins]] ([[Anions|anions]]) will bind to a positively charged diethylaminoethylcellulose (DEAE-cellulose) columns. The negative proteins bound to column can by eluted by adding a negatively charged [[Buffer|buffer]] which will compete with the [[Protein|protein]] for binding to the column. Proteins that have lower charge density will be eluted first from the column <ref>Berg J., Tymoczko J and Stryer L. (2007) Biochemistry, 6th edition, New York: WH Freeman</ref>. | For example negatively charged [[Proteins|proteins]] ([[Anions|anions]]) will bind to a positively charged [[diethylaminoethylcellulose |diethylaminoethylcellulose ]](DEAE-cellulose) columns. The negative proteins bound to column can by eluted by adding a negatively charged [[Buffer|buffer]] which will compete with the [[Protein|protein]] for binding to the column. Proteins that have lower charge density will be eluted first from the column <ref>Berg J., Tymoczko J and Stryer L. (2007) Biochemistry, 6th edition, New York: WH Freeman</ref>. | ||
The stationary phase is an insoluble matrix which will either carry a positve or negative charge. Whilst diethylaminoethylcellulose (DEAE-cellulose) is positively charged, an negatively charged matrix includes carboxymethylcellulose (CM-cellulose) | The stationary phase is an insoluble matrix which will either carry a positve or negative charge. Whilst diethylaminoethylcellulose (DEAE-cellulose) is positively charged, an negatively charged matrix includes [[carboxymethylcellulose|carboxymethylcellulose]] (CM-cellulose) <ref>Alberts et al. (2008) Molecular Biology Of The Cell, 5th Edition. Chapter 8, page 514.</ref>.<br> | ||
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=== References === | === References === | ||
Revision as of 17:25, 23 October 2012
Ion-exchange chromatography separates proteins according to their net charge. The stationary phase of the column will have a specific charge.
For example negatively charged proteins (anions) will bind to a positively charged diethylaminoethylcellulose (DEAE-cellulose) columns. The negative proteins bound to column can by eluted by adding a negatively charged buffer which will compete with the protein for binding to the column. Proteins that have lower charge density will be eluted first from the column [1].
The stationary phase is an insoluble matrix which will either carry a positve or negative charge. Whilst diethylaminoethylcellulose (DEAE-cellulose) is positively charged, an negatively charged matrix includes carboxymethylcellulose (CM-cellulose) [2].
References