title: Use of monoclonal antibodies for expression cloning.
abstract: This unit details the use of transient expression in mammalian cells to screen cDNA libraries with monoclonal antibodies (MAb) to isolate cDNA clones encoding cell-surface and intracellular proteins. The first protocol in this unit describes the cloning of cDNAs encoding cell-surface antigens. Several steps in this protocol involve transfection procedures that are described in greater detail elsewhere in this volume. The second protocol is a modification that facilitates isolation of cDNAs encoding antigens that are expressed intracellularly. Both protocols are designed for use with the expression vector CDM8, which contains a polylinker for subcloning double-stranded cDNA.
authors.affiliation: Bristol-Myers Squibb, Seattle, Washington, USA.
authors.forename: Diane
authors.initials: D
authors.lastname: Hollenbaugh
authors.forename: Alejandro
authors.initials: A
authors.lastname: Aruffo
authors.forename: Bryan
authors.initials: B
authors.lastname: Jones
authors.forename: Peter
authors.initials: P
authors.lastname: Linsley
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000235:genetics* / Q000276:immunology; D000954:Antigens, Surface / Q000235:genetics* / Q000276:immunology; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D003637:DEAE-Dextran; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics; D007163:Immunosorbent Techniques / Q000295:instrumentation; D042541:Intracellular Space; D008322:Mammals; D011143:Polyvinyl Chloride / Q000031:analogs & derivatives; D013104:Spheroplasts / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D000954:Antigens, Surface; D018076:DNA, Complementary; C029547:polyvinylidene chloride; D011143:Polyvinyl Chloride; D003637:DEAE-Dextran