Multiple Epitope Tagging of Expressed Proteins for Enhanced Detection | Zendy

Ron Hernan | Zendy; Ken Heuermann | Zendy; Bill Brizzard | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Multiple Epitope Tagging of Expressed Proteins for Enhanced Detection

Author(s) -

Ron Hernan,

Ken Heuermann,

Bill Brizzard

Publication year - 2000

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/00284pf01

Subject(s) - fusion protein , epitope , western blot , microbiology and biotechnology , alkaline phosphatase , expression vector , escherichia coli , flag tag , biology , vector (molecular biology) , blot , affinity chromatography , flag (linear algebra) , biochemistry , recombinant dna , gene , antigen , enzyme , genetics , algebra over a field , mathematics , pure mathematics

Three FLAG epitopes have been incorporated into the mammalian expression vector pCMV-5 to create a transient expression vector, p3XFLAG-CMV-7. The vector was designed to express FLAG fusion proteins that can be detected at tenfold lower expression levels than the current FLAG fusion protein expression system. The usefulness of this expression and detection system was demonstrated by expression of bacterial alkaline phosphatase in COS-7 cells. In addition, 3XFLAG bacterial alkaline phosphatase was expressed in Escherichia coli, purified on anti-FLAG M2 affinity gel, and detection of 500 pg of purified protein by Western blot analysis is demonstrated.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research