Summary
Development of immunoblots is commonly performed using enzyme-labeled antibodies, which convert soluble substrates into insoluble colored products. A simple, rapid, and sensitive alternative method that produces low background and allows a rapid quantitative evaluation is the use of radiolabeled antibodies or protein A conjugates. Here, we describe the use of iodinated secondary antibodies for immunodetection of an autoantigen during HPLC purification.
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References
Weser, S., Bachmann, M., Seifart, K.H., and Meissner, W. (2000) Transcription efficiency of human polymerase III genes in vitro does not depend on the RNP-forming autoantigen La. Nucleic Acids Res. 15, 3935–3942.
Tröster, H.,Bartsch,H.,Klein,R.,Metzger,T.E.,Pollak,G., et al (1995) Activation of a murautoreactive B cell by immunization with human recombinant autoantigen La/SS-B: characterization of the autoepitope. J Autoimmun. 8, 825–842.
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Acknowledgements
Special thanks for excellent technical support to K. Joanne and Tim Gross (Oklahoma Medical Research Foundation, Oklahoma City, USA).
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Bartsch, H., Franke, C., Bachmann, M. (2009). Immunoblotting using Radiolabeled Reagents for Detection. In: Kurien, B., Scofield, R. (eds) Protein Blotting and Detection. Methods in Molecular Biology, vol 536. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-542-8_45
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DOI: https://doi.org/10.1007/978-1-59745-542-8_45
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Publisher Name: Humana Press, Totowa, NJ
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