Rat liver fixed by perfusion with low glutaraldehyde concentrations was incubated in diaminobenzidine-containing medium to stain for peroxidase. Endogenous peroxidatic activity was found not only in Kupffer cells but also in the endothelial cells lining the sinusoids and central veins. The reaction product was localized in the nuclear envelope and endoplasmic reticulum. The peroxidatic activity in endothelial cells showed a concentration-dependent sensitivity to glutaraldehyde: in liver samples fixed with 0.25% glutaraldehyde, approx. 23% of the sinusoidal endothelial cells and 65% of central vein endothelium were peroxidase-positive; with 0.5% glutaraldehyde, only approx. 8% of the sinusoidal endothelial cells contained detectable amounts of the reaction product; with 1.5% glutaraldehyde all endothelial cells were consistently peroxidase-negative. No peroxidatic activity could be found in liver endothelial cells following isolation by centrifugal elutriation. Endothelial cell peroxidase may possibly be involved in defense responses of liver and/or, as a part of prostaglandin synthase system, in prostanoid production.