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Biotechniques. 1996 Jan;20(1):136-41.

Vectors for expression and secretion of FLAG epitope-tagged proteins in mammalian cells.

Author information

1
Scientific Imaging Systems, Eastman Kodak Company, New Haven, CT, USA.

Abstract

The FLAG peptide, AspTyrLysAspAspAspAspLys, has been used as an epitope tag in a variety of cell types. The modification of the cytomegalovirus (CMV) promoter containing vector, pCMV5, to create two transient expression vectors designed for secretion and intracellular expression of FLAG-fusion proteins in mammalian cells is described. As a functional test, the bacterial alkaline phosphatase gene was cloned into both vectors, and anti-FLAG monoclonal antibodies were used for detection of FLAG epitope-tagged bacterial alkaline phosphatase in mammalian cells. In addition, secreted bacterial alkaline phosphatase was purified from the extracellular medium by anti-FLAG affinity chromatography.

PMID:
8770418
DOI:
10.2144/96201pf01
[Indexed for MEDLINE]
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