Affinity purification of monospecific antibodies from polyclonal sera as a means for the identification of differentially expressed genes and proteins.
The identification and characterization of genes and proteins that are either cell-type specific or exhibit an otherwise spatially or temporally restricted expression pattern is a crucial step toward understanding the complex interactions between the different cell types in an organism. We demonstrate here that monospecific polyclonal antibodies purified from rabbit antisera by a simple Western blotting and reelution technique can be successfully used for the detection of such proteins, as well as the corresponding genes by screening cDNA expression libraries. This strategy is discussed as a supplement to other approaches, such as the generation of tissue-specific monoclonal antibodies and the "subtractive hybridization technique."
von Boxberg
Y
Y
Max-Planck-Institut für Entwicklungsbiologie, Tübingen, Germany.
Kemmner
W A
WA
D000818:Animals; D000906:Antibodies / Q000097:blood / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000097:blood / Q000302:isolation & purification*; D015153:Blotting, Western / Q000379:methods; D002642:Chick Embryo; D002846:Chromatography, Affinity / Q000379:methods; D018076:DNA, Complementary / Q000096:biosynthesis; D015870:Gene Expression; D015723:Gene Library; D005821:Genetic Techniques; D007150:Immunohistochemistry / Q000379:methods; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis*; D011817:Rabbits / Q000276:immunology; D012160:Retina / Q000166:cytology / Q000196:embryology / Q000378:metabolism*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011506:Proteins