Abstract
Expression of recombinant proteins is a standard technique in molecular biology and a wide variety of prokaryotic as well as eukaryotic expression systems are currently in use. A limiting step is often the purification of the expressed recombinant protein, particularly if mammalian expression systems that yield low expression levels are employed. Here, we discuss the advantages and restrictions of tagging recombinant proteins with histidines and purifying them by Ni2+-NTA chromatography.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- GST:
-
glutathione S-transferase
- NTA:
-
nitrilotriacetic acid
- His:
-
histidine
- PAGE:
-
polyacrylamide gel electrophoresis
References
Uhlen M & Moks T (1990) Methods Enzymology 185: 129–143
Smith DB & Johnson KS (1988) Gene 67: 31–40
Moks T, Abrahmsen L, Oesterlöf B, Josephson S, Oestling M, Enfors SO, Persson I, Nilsson B & Uhlen M (1987) Bio/Technology 5: 379–382
Hopp TP, Prickett KS, Price VL, Libby RT, March CJ, Ceretti DP, Urdall DL & Conlon PJ (1988) Bio/Technology 6: 1204–1210
Hochuli E, Döbeli H & Schacher A (1987) J. Chromatography 411: 177–184
Porath J, Carlsson J, Olsson I & Belfrage G (1975) Nature 258: 598–599
Hochuli E, Bannwarth W, Döbeli H, Gentz R & Stüber D (1988) Bio/Technology 6: 1321–1325
Janknecht R, De Martynoff G, Lou J, Hipskind RA, Nordheim A & Stunnenberg HG (1991) Proc. Natl. Acad. Sci. USA 88: 8972–8976
Janknecht R & Nordheim A (1992) Gene 121: 321–324
Nikolow DB, Hu SH, Lin J, Gasch A, Hoffmann A, Horikoshi M, Chua NH, Roeder RG & Burley SK (1992) Nature 360: 40–46
Kozak M (1984) Nucleic Acids Res. 12: 857–872
Schmitt J, Pohl J & Stunnenberg HG (1993) Gene, in press
Schmitt J & Stunnenberg HG (1993) Nucleic Acids Res. 21: 2673–2681
Stunnenberg HG (1993) BioEssays 15: 309–315
Bugge TH, Pohl J, Lonnoy O & Stunnenberg HG (1992) EMBO J. 11: 1409–1418
Suzuki N, Rohdewohld H, Neuman T, Gruss P & Schöler HR (1990) EMBO J. 9: 3723–3732
Arnold FH (1991) Bio/Techechnology 9: 151–156
Hoffmann A & Roeder RG (1991) Nucleic Acids Res. 19: 6337–6338
Brewer SJ & Sassenfeld HM (1989) In: Harris ELV & Angal S (Eds) Engineering proteins for purification (pp: 91–111) IRL Press, Oxford
Studier FW & Moffat BA (1986) J. Mol. Biol. 189: 113–130
Janknecht R, Hipskind RA, Houthaeve T, Nordheim A & Stunnenberg HG (1992) EMBO J. 11: 1045–1054
Dignam JD, Lebowitz RM & Roeder RG (1983) Nucleic Acids Res. 11: 1475–1489
Gounari F, De Francesco R, Schmitt J, van der Vliet PC, Cortese R & Stunnenberg H (1990) EMBO J. 9: 559–566
Harlow E & Lane D (1988) Antibodies. A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Schöler HR, Balling R, Hatzopoulos AK, Suzuki N & Gruss P (1989) EMBO J. 8: 2551–2557
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schmitt, J., Hess, H. & Stunnenberg, H.G. Affinity purification of histidine-tagged proteins. Mol Biol Rep 18, 223–230 (1993). https://doi.org/10.1007/BF01674434
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01674434