Protein Precipitation Using Ammonium Sulfate

Paul T. Wingfield1

1 National Institutes of Health, Bethesda, Maryland
Publication Name:  Current Protocols in Protein Science
Unit Number:  Appendix 3F
DOI:  10.1002/0471140864.psa03fs84
Online Posting Date:  April, 2016
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The basic theory of protein precipitation by addition of ammonium sulfate is presented, and the most common applications are listed. Tables are provided for calculating the appropriate amount of ammonium sulfate to add to a particular protein solution. © 2016 by John Wiley & Sons, Inc.

Keywords: ammonium sulfate; ammonium sulfate tables; protein concentration; protein purification

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Table of Contents

  • Basic Theory
  • Tips and Guidelines
  • Common Applications
  • Ammonium Sulfate Tables
  • Tables
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Literature Cited

Literature Cited
   Dawson, R.M.C. , Elliot, D.C. , Elliot, W.H. , and Jones, K.M. 1986. Data for Biochemical Research (3rd ed.), p. 537. Oxford Science Publications, Clarendon Press, Oxford.
   Green, A.A. and Hughes, W.L. 1955. Protein solubility on the basis of solubility in aqueous solutions of salts and organic solvents. Methods Enzymol. 1:67‐90. doi: 10.1016/0076‐6879(55)01014‐8.
   Hagel, L. 1998. Gel‐filtration chromatography. Curr. Protoc. Protein Sci. 14:8.3.1‐8.3.30.
   Jakoby, W.B. 1971. Crystallization as a purification technique. Methods Enzymol. 22:248‐252. doi: 10.1016/0076‐6879(71)22025‐5.
   Kennedy, R. M. 1995. Hydrophobic‐interaction chromatography. Curr. Protoc. Protein Sci. 00:8.4.1‐8.4.21.
   Kita, Y. , Arakawa, T. , Lin, T.‐Y. , and Timasheff, S. 1994. Contribution of the surface free energy perturbations to protein‐solvent interactions. Biochemistry 33:1517‐1589. doi: 10.1021/bi00254a029.
   Lovrien, R.E. and Matulis, D. 1997. Selective precipitation of proteins. Curr. Protoc. Protein Sci. 7:4.5.1‐4.5.36.
   Mitchinson, C. and Pain, R.H. 1986. The effect of sulphate and urea on the stability and reversible unfolding of β‐lactamase from Staphylococcus aureus . J. Mol. Biol. 184:331‐342. doi: 10.1016/0022‐2836(85)90384‐5.
   Parsegian, V.A. 1995. Hopes for Hofmeister. Nature 378:335‐336. doi: 10.1038/378335a0.
   Rupley, J.A. , Gratton, E. , and Careri, G. 1983. Water and globular proteins. Trends Biochem. Sci. 8:18‐22. doi: 10.1016/0968‐0004(83)90063‐4.
   Schagger, H. 1994. Chromatographic techniques and basic operations in membrane protein purification. In A Practical Guide to Membrane Protein Purification (G. von Jagow and H. Schagger, eds.) pp. 23‐57. Academic Press, San Diego.
   Timasheff, S.N. and Arakawa, T. 1997. Membrane protein purification stabilization of protein structure by solvents. In Protein Structure: A Practical Approach. 2nd ed. ( T.E., Creighton , ed.) pp. 349‐364. IRL Press at Oxford University Press, Oxford.
   Wingfield, P.T. , Stahl, S.J. , Payton, M.A. , Vankatesan, S. , Misra, M. , and Steven, A.C. 1991. HIV‐1 Rev expressed in recombinant Escherichia coli: Purification, polymerization and conformational properties. Biochemistry 30:7527‐7534. doi: 10.1021/bi00244a023.
   Wingfield, P.T. 2014. Preparation of soluble proteins from Escherichia coli . Curr. Protoc. Protein Sci. 78:6.2.1‐6.2.22. doi: 10.1002/0471140864.ps0602s78.
   Wood, W.I. 1976. Tables for the preparation of ammonium sulfate solutions. Anal. Biochem. 73:250‐257. doi: 10.1016/0003‐2697(76)90165‐2.
Key Reference
   Wood, 1976. See above.
  Tables A.3F.2 to A.3F.5 are reprinted from this publication.
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