Intensive immunofluorescence staining methods for low expression protein: Detection of intestinal stem cell marker LGR5

Masaki Yamazaki, Atsuhiko Kato, Yoko Zaitsu, Takeshi Watanabe, Makoto Iimori, Shinichi Funahashi, Hiroyuki Kitao, Hiroshi Saeki, Eiji Oki, Masami Suzuki

Research output: Contribution to journalComment/debate

3 Citations (Scopus)

Abstract

Leucine-rich repeat-containing G-protein coupled receptor 5, or LGR5, is a molecule that recognizes stem cells in multiple organs and also in colon cancer. Previously, we have developed monoclonal antibodies specific to LGR5 protein that can be used for immunofluorescence staining, but because a very low level of LGR5 protein is expressed, the visualization technique needed to be enhanced. To develop procedures to detect LGR5 protein in various specimens by immunofluorescence staining, we evaluated the Alexalabeled streptavidin biotin (LSAB), the Qdot, and the tyramide methods. The detection sensitivity was highest in the tyramide method followed by the Qdot method, whereas subcellular localization of the protein was most clear in the Qdot method, because the Qdot method gave a high S/N ratio that could show a low background. Thus, the tyramide method is superior to the Q-dot method for intensifying the signal of a low expression protein, and the Qdot method is superior to the tyramide method for identifying the subcellular localization of the target protein. The results of the present study will be helpful in providing more insight into the pathophysiological roles of LGR5-positive cancer stem cells and in developing therapeutic approaches for targeting cancer stem cells.

Original languageEnglish
Pages (from-to)159-164
Number of pages6
JournalActa Histochemica et Cytochemica
Volume48
Issue number5
DOIs
Publication statusPublished - Oct 29 2015

Fingerprint

Stem cells
Fluorescent Antibody Technique
Stem Cells
Staining and Labeling
Proteins
Neoplastic Stem Cells
Streptavidin
Biotin
G-Protein-Coupled Receptors
Leucine
Visualization
Monoclonal Antibodies
Molecules
Colonic Neoplasms

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Biochemistry
  • Physiology
  • Histology
  • Cell Biology

Cite this

Intensive immunofluorescence staining methods for low expression protein : Detection of intestinal stem cell marker LGR5. / Yamazaki, Masaki; Kato, Atsuhiko; Zaitsu, Yoko; Watanabe, Takeshi; Iimori, Makoto; Funahashi, Shinichi; Kitao, Hiroyuki; Saeki, Hiroshi; Oki, Eiji; Suzuki, Masami.

In: Acta Histochemica et Cytochemica, Vol. 48, No. 5, 29.10.2015, p. 159-164.

Research output: Contribution to journalComment/debate

Yamazaki, Masaki ; Kato, Atsuhiko ; Zaitsu, Yoko ; Watanabe, Takeshi ; Iimori, Makoto ; Funahashi, Shinichi ; Kitao, Hiroyuki ; Saeki, Hiroshi ; Oki, Eiji ; Suzuki, Masami. / Intensive immunofluorescence staining methods for low expression protein : Detection of intestinal stem cell marker LGR5. In: Acta Histochemica et Cytochemica. 2015 ; Vol. 48, No. 5. pp. 159-164.
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AU - Iimori, Makoto

AU - Funahashi, Shinichi

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