Abstract
Highly specific antibodies directed against minor proteins, present in small amounts in biological fluids, or against nonsoluble cytoplasmic or membraneous proteins, are often difficult to obtain. The main reasons for this are the small amounts of protein available after the various classical purification processes and the low purity of the proteins.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Knudsen, K. A. (1985) Proteins transferred to nitrocellulose for use as immunogens. Anal. Biochem. 147, 285–288.
O’Farrel, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021.
O’Farrel, P. Z., Goodman, H. M., and O’Farrel, P. H. (1977) High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12, 1133–1142.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685.
Burnette, W. N. (1981) Electrophoretic transfer of proteins from sodium dodecylsulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal. Biochem. 112, 195–203.
Vaitukaitis, J., Robbins, J. B., Nieschlag, E., and Ross, G. T. (1971) A method for producing specific antisera with small doses of immunogen. J. Clin. Endocrinol. 33, 980–988.
Dunn, S. D. (1986) Effects of the modification of transfer buffer composition and the renaturation of proteins in gels on the recognition of proteins on Western blots by monoclonal antibodies. Anal. Biochem. 157, 144–153
Sigel, M. B., Sinha, Y. N., and Vanderlaan, W. P. (1983) Production of antibodies by inoculation into lymph nodes. Methods Enzymol. 93, 3–12.
Preziosi, L., Michel, G. P. F., and Baratti, J. (1990) Characterisation of sucrose hydrolising enzymes of Zymomonas mobilis. Arch. Microbiol. 153, 181–186.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1992 The Humana Press, Inc., Totowa, NJ
About this protocol
Cite this protocol
Diano, M., Bivic, A.L., Hirn, M. (1992). Raising Polyclonal Antibodies Using Nitrocellulose-Bound Antigen. In: Manson, M.M. (eds) Immunochemical Protocols. Methods in Molecular Biology, vol 10. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-204-3:13
Download citation
DOI: https://doi.org/10.1385/0-89603-204-3:13
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-204-0
Online ISBN: 978-1-59259-497-9
eBook Packages: Springer Protocols