The assay for NADH-ferrihemoglobin reductase (NADH-FR) was optimized for avian blood samples. In this assay the pH optimum for Japanese quail red cell NADH-FR was 5.5, which was close to the enzyme's pI. Enzyme kinetic parameters were determined for quail, chicken and turkey NADH-FR. Preparation of erythrocyte ghost-cells and subsequent fractionation showed that the enzyme was present in the plasma membrane as well as in the nuclear membrane, while Triton X-100 treatment gave a release of enzyme activity from the membrane. In the cytosolar fraction of avian red cells no NADH-FR could be detected.