Immunoprecipitation Techniques

Abstract

Immunoprecipitations are based on two types of interaction: between antigen and antibody, and between antibody and an immunoadsorbent. The antigen–antibody complex will be bound to an insoluble matrix that can be pelleted and removed from the bulk protein. Immunoprecipitation allows rapid and efficient purification of proteins in a native and biochemically active form.

Keywords: antibody; antigen; epitope; immunoprecipitation; protein A

Figure 1.

Flow diagram of general procedure.

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References

Ausubel FA, Brent R, Kingston RE et al. (eds) (1998) Current Protocols in Molecular Biology. New York: Wiley.

Getzoff ED, Geysen HM, Rodda SJ et al. (1987) Mechanisms of antibody binding to a protein. Science 235: 1191–1196.

Grussenmeyer T, Scheidtmann KH, Hutchinson MA, Eckhart W and Walter G (1985) Complexes of polyoma virus medium T antigen and cellular proteins. Proceedings of the National Academy of Sciences of the USA 82: 7952–7954.

Harlow E and Lane DW (1988) Antibodies, A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

Kapoor M and Lozano G (1998) Functional activation of p53 via phosphorylation following DNA damage by UV but not γ radiation. Proceedings of the National Academy of Sciences of the USA 95: 2834–2837.

McKay RDG (1981) Binding of SV40 T antigen related protein to DNA. Journal of Molecular Biology 145: 471–488.

Milner J (1995) Flexibility: the key to p53 function? Trends in Biochemical Sciences 20: 49–51.

Mueller E, Boldyreff B and Scheidtmann KH (1993) Characterization of protein kinase activities associated with p53‐large‐T immune complexes from SV40‐transformed rat cells. Oncogene 8: 2193–2205.

Scheidtmann KH, Reinartz S and Schlebusch H (1997) Immunological detection of proteins of known sequence. In Creighton TE (ed.) Protein Structure, pp. 59–89. Oxford: IRL Press.

Schickedanz J, Scheidtmann KH and Walter G (1986) Kinetics of nuclear transport and oligomerization of simian virus 40 large T antigen. Virology 148: 47–57.

Shiio Y, Itoh M and Inoue J (1995) Epitope tagging. Methods in Enzymology 254: 497–502.

Simanis V and Lane DP (1985) An immunoaffinity purification procedure for simian virus 40 T antigen. Virology 144: 88–100.

Staufenbiel M and Deppert W (1983) Different structural systems of the nucleus are targets for SV40 large T antigen. Cell 33: 173–181.

Walter G (1986) Production and use of antibodies against synthetic peptides. Journal of Immunological Methods 88: 149–161.

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How to Cite close
Scheidtmann, Karl‐Heinz(Apr 2001) Immunoprecipitation Techniques. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0001180]