Use of Immunomatrix Methods to Improve Protein-Protein Interaction Detection
Immunoprecipitation (IP) and coimmunoprecipitation (co-IP) are key techniques for studying protein-protein interactions. These methods utilize immobilized protein A or protein G to isolate antibody-bound target antigens. The main disadvantage of traditional immunoprecipitation and coimmunoprecipitat...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Hindawi Limited
2003-01-01
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| Series: | Journal of Biomedicine and Biotechnology |
| Online Access: | http://dx.doi.org/10.1155/S1110724303209256 |
| Summary: | Immunoprecipitation (IP) and coimmunoprecipitation (co-IP) are key
techniques for studying protein-protein interactions. These
methods utilize immobilized protein A or protein G to isolate
antibody-bound target antigens. The main disadvantage of
traditional immunoprecipitation and coimmunoprecipitation is that
the conditions used to elute the precipitated antigen also
release the antibody, contaminating the antigen and destroying
the antibody support. To overcome these problems, we describe two
methods to generate a reusable antibody support by cross-linking
the antibody to immobilized protein A or protein G, or by
coupling it directly to the resin. Our
studies have demonstrated that the immobilization efficiency for
the antibody coupling method was similar for several species of
antibody. Furthermore, we illustrate that using both methods of
antibody immobilization yields IP and co-IP results similar to
traditional protocols but eliminate the antibody heavy and light
chains contamination. |
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| ISSN: | 1110-7243 1110-7251 |