Use of monoclonal antibodies for expression cloning.
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.
Hollenbaugh
Diane
D
Bristol-Myers Squibb, Seattle, Washington, USA.
Aruffo
Alejandro
A
Jones
Bryan
B
Linsley
Peter
P
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
D000911:Antibodies, Monoclonal; D000941:Antigens; D000954:Antigens, Surface; D018076:DNA, Complementary; C029547:polyvinylidene chloride; D011143:Polyvinyl Chloride; D003637:DEAE-Dextran