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 Citation (Scopus)

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

Fingerprint

Transferrin Receptors
K562 Cells
Transferrin
Rosette Formation
Erythrocytes
Amino Acid Sequence Homology
Erythroid Cells
Lactoferrin
Density Gradient Centrifugation
Cell Separation
Centrifugation
Leukemia
Monoclonal Antibodies
Cell Line

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Visualization of the receptor for transferrin on K562 cells by a rosette-forming assay. / Yamada, Yasuaki; Jacobsen, Donald W.; Green, Ralph.

In: Journal of Immunological Methods, Vol. 86, No. 1, 22.01.1986, p. 95-101.

Research output: Contribution to journalArticle

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