Simultaneous detection of double-stranded RNA-induced protein kinase and its specific mRNA by single cell analysis

Biotechnol Lett. 2005 Jun;27(12):875-80. doi: 10.1007/s10529-005-6725-3.

Abstract

Fluorescent in situ hybridization was combined with flow cytometry to detect the expression of the double-stranded-RNA-induced protein kinase (PKR) in single cells. Labeled anti-sense oligonucleotide was used to target the specific mRNA while the protein was targeted with an antibody. It was demonstrated that the PKR-mRNA signal could be protected through a lengthy immunostaining procedure. The expression pattern of the PKR-mRNA with respect to DNA content was shown to be comparable to that of 18S ribosomal RNA.

MeSH terms

  • Cell Line, Tumor
  • Flow Cytometry / methods
  • Fluorescent Dyes
  • Humans
  • In Situ Hybridization, Fluorescence / methods
  • Protein Kinases / genetics*
  • Protein Kinases / metabolism
  • RNA, Double-Stranded / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Reproducibility of Results

Substances

  • Fluorescent Dyes
  • RNA, Double-Stranded
  • RNA, Messenger
  • Protein Kinases