Construction of multiple-epitope tag sequence by PCR for sensitive Western blot analysis
- 1 June 1997
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 25 (11) , 2231-2232
- https://doi.org/10.1093/nar/25.11.2231
Abstract
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.Keywords
This publication has 6 references indexed in Scilit:
- [33] Epitope taggingPublished by Elsevier ,1995
- High-level expression, purification, and enzymatic characterization of full-length Thermus aquaticus DNA polymerase and a truncated form deficient in 5' to 3' exonuclease activity.Genome Research, 1993
- Association of Sos Ras exchange protein with Grb2 is implicated in tyrosine kinase signal transduction and transformationNature, 1993
- Vaccinia Virus Encodes a Family of Genes with Homology to Serine Proteinase InhibitorsJournal of General Virology, 1989
- At least six nucleotides preceding the AUG initiator codon enhance translation in mammalian cellsJournal of Molecular Biology, 1987
- Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.Molecular and Cellular Biology, 1985