Abstract
Immunoprobing for specific peptides on solid supports such as nitrocellulose requires prior blocking of nonspecific protein binding sites. Many proteins, or protein mixtures, have been used for this, including bovine serum albumin, gelatin, and calf serum, but the cheapest and most efficient method makes use of dried milk protein.
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References
Johnson, D. A., Gautsch, J.W., Sportsman, J.R., and Elder, J.H. (1984) Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal. Tech. 1, 3–8.
Hum, A.L. and Chantler, S.M. (1980) Production of reagent antibodies. Methods in Enzymol. 70, 104–142.
Austin, S.A., Pollard, J.W., Jagus, R., and Clemens, M. (1986) Regulation of polypeptide initiation and activity of initiation factor 2 in Chinese hamster ovary cell mutants containing temperature-sensitive aminoacyl tRNA synthetases. Eur. J. Biochem. 157, 39–47.
Brada, D. and Roth, J. (1984) Golden blot. Detection of polyclonal and monoclonal antibodies bound to antigens on nitrocellulose by protein A-gold complexes. Anal Biochem. 142, 79–83.
Burnette, W.N. (1981) Western blotting. Anal. Biochem. 112, 195–203.
Miskimins, W.K., Roberts, M.P., McClelland, A., and Ruddle, F.H. (1985) Proc. Natl. Acad. Sci. 82, 6741–6744.
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© 1988 The Humana Press Inc.
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Jagus, R., Pollard, J.W. (1988). Use of Dried Milk for Immunoblotting. In: Walker, J.M. (eds) New Protein Techniques. Methods in Molecular Biology™, vol 3. Humana Press. https://doi.org/10.1385/0-89603-126-8:403
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DOI: https://doi.org/10.1385/0-89603-126-8:403
Publisher Name: Humana Press
Print ISBN: 978-0-89603-126-5
Online ISBN: 978-1-59259-490-0
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