Visualization of the receptor for transferrin on K562 cells by a rosette-forming assay

Yasuaki Yamada, Donald W. Jacobsen, Ralph Green

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

A method is described to visualize the receptor for transferrin by a rosette-forming assay. K562 cells, a human pre-erythroid leukemia cell line, were used as a source of receptor-positive cells. Bovine erythrocytes coated with human transferrin by the CrCl3 method were used as indicator cells. Rosette formation occurred at 37°C but not at 4°C, and was inhibited by soluble transferrin in a dose-dependent manner. Human lactoferrin, which is known to have considerable amino acid sequence homology with transferrin, did not inhibit rosette formation with transferrin-coated bovine erythrocytes. A variety of blocking and non-blocking monoclonal antibodies against the human transferrin receptor (42/6, B3/25 and OKT9) inhibited rosette formation. Rosettes are able to withstand low-speed centrifugation (130 × g, 2 min) making this method potentially useful for cytochemical identification of receptor-positive or receptor-negative cells. Also, it was possible to enrich receptor-positive cells by separation of rosette-forming from non-rosette-forming cells using a density gradient centrifugation technique.

Original languageEnglish (US)
Pages (from-to)95-101
Number of pages7
JournalJournal of Immunological Methods
Volume86
Issue number1
DOIs
StatePublished - Jan 22 1986
Externally publishedYes

Keywords

  • Anti-transferrin receptor antibody
  • K562 cells
  • Rosette-forming assay
  • Surface marker
  • Transferrin receptor

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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