Vectors for Expression and Secretion of FLAG Epitope-Tagged Proteins in Mammalian Cells | Zendy

Richard Chubet | Zendy; Bill Brizzard | Zendy

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Vectors for Expression and Secretion of FLAG Epitope-Tagged Proteins in Mammalian Cells

Author(s) -

Richard Chubet,

Bill Brizzard

Publication year - 1996

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/96201pf01

Subject(s) - epitope , monoclonal antibody , biology , alkaline phosphatase , microbiology and biotechnology , flag (linear algebra) , expression vector , secretion , fusion protein , extracellular , biochemistry , gene , antibody , enzyme , recombinant dna , genetics , algebra over a field , mathematics , pure mathematics

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.

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