Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing

Zsuzsanna E. Toth, Tal Shahar, Ronen Leker, Ildiko Szalayova, András Bratincsák, Sharon Key, Anna Lonyai, Krisztián Németh, Éva Mezey

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

The green fluorescent protein (GFP) is among the most commonly used expression markers in biology. GFP-tagged cells have played a particularly important role in studies of cell lineage. Sensitive detection of GFP is crucially important for such studies to be successful, and problems with detection may account for discrepancies in the literature regarding the possible fate choices of stem cells. Here we describe a very sensitive technique for visualization of GFP. Using it we can detect about 90% of cells of donor origin while we could only see about 50% of these cells when we employ the methods that are in general use in other laboratories. In addition, we provide evidence that some cells permanently silence GFP expression. In the case of the progeny of bone marrow stem cells, it appears that the more distantly related they are to their precursors, the more likely it is that they will turn off the lineage marker.

Original languageEnglish
Pages (from-to)1943-1950
Number of pages8
JournalExperimental Cell Research
Volume313
Issue number9
DOIs
StatePublished - 15 May 2007
Externally publishedYes

Keywords

  • Cell fate
  • Green fluorescent protein
  • Hematopoetic stem cell
  • Immunohistochemistry
  • Lineage tracking
  • Stem cell transplantation

ASJC Scopus subject areas

  • Cell Biology

Fingerprint

Dive into the research topics of 'Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing'. Together they form a unique fingerprint.

Cite this