Purification of recombinant proteins and study of protein interaction by epitope tagging.
A protein molecule can be engineered to include a short stretch of residues corresponding to an epitope to facilitate its subsequent biochemical and immunological analysis; a technique often referred to as "epitope tagging." This unit presents a protocol for small-scale immunoprecipitation of epitope-tagged recombinant proteins expressed in transiently transfected mammalian cells. The immunoprecipitant can then be analyzed by SDS-PAGE. An immunoprecipitation protocol is also provided that has been optimized for use with a baculovirus overexpression system. An Alternate Protocol describes how multisubunit complexes can be assembled by starting with a core protein affixed to beads via an epitope tag, and adding the other members of the complex in a stepwise manner.
Zhang
N
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Tularik, Inc., South San Francisco, California, USA.
Chen
J L
JL
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001665:Binding Sites; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005822:Genetic Vectors; D006801:Humans; D047468:Immunoprecipitation; D011485:Protein Binding; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D014162:Transfection
D000939:Epitopes; D011994:Recombinant Proteins