RVV test. Aliquots (150 μL) of citrated platelet-poor plasma were mixed with 20 μL of PBS containing increasing numbers of normal (•) or band 3 null (○) RBCs from adult mice. After incubation at 37°C for 2 minutes, 150 μL of RVV test solution containing RVV and calcium was added. The time required for the formation of a fibrin clot was measured with a fibrometer. Note that RBCs from normal mice have no effect on the RVV clotting time, whereas RBCs from band 3 null mice significantly shorten the RVV clotting time. The values shown are the mean of four independent experiments ± standard deviation (SD).