Skip to main content
Fig. 1 | Genes and Environment

Fig. 1

From: Standard protocol for the total red blood cell Pig-a assay used in the interlaboratory trial organized by the Mammalian Mutagenicity Study Group of the Japanese Environmental Mutagen Society

Fig. 1

Principle of the Pig-a assay and flow cytometry analysis. a Pig-a is an essential gene for synthesis of the glycosylphosphatidylinositol (GPI) anchor. In wild-type cells, GPI anchors and CD59, a GPI-anchored protein marker, are synthesized independently and GPI tethers CD59 to the cell surface. However, in Pig-a mutant cells, CD59 proteins on the cell surface are reduced because GPI anchors are not synthesized due to Pig-a gene mutation(s). Thus, Pig-a mutant cells do not react with FITC-conjugated anti-CD59 antibodies while wild-type cells react to the antibodies and fluoresce. b Peripheral blood is stained with fluorescent-labeled antibodies. Cells are gated by light scatter and then analyzed by flow cytometry for HIS49 rat erythroid marker expression. HIS49-positive cells are further analyzed for CD59 expression and Pig-a mutant cells are detected as the FITC-negative population

Back to article page

Genes and Environment

ISSN: 1880-7062

Contact us