Construction of multiple-epitope tag sequence by PCR for sensitive Western blot analysis | Zendy

Kazunori Nakajima | Zendy

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Construction of multiple-epitope tag sequence by PCR for sensitive Western blot analysis

Author(s) -

Kazunori Nakajima

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.11.2231

Subject(s) - epitope , biology , microbiology and biotechnology , western blot , complementary dna , virology , linear epitope , antibody , genetics , gene

Epitope tagging is a powerful technique to characterize a recombinantly expressed protein encoded by cDNA without the purification of the protein and the immunization of animals. In some cases, however, the expression of a tagged protein is too low to analyze by Western blot. We have developed a simple method to generate tandem repetitive nucleotide sequence by PCR, which allows us to label a protein of interest with a multiple-epitope tag. When five myc epitopes were attached to vaccinia virus protein CrmA, its signal was multiplied 5.8 times in Western blot analysis, compared with that of one epitope-tagged CrmA.

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