Rye and wheat bran are excellent natural sources of plant sterols in the diet. Their content, however, may vary according to processing. Thermal (roasting and heating in a microwave oven), mechanical (milling and cryogenic grinding), and enzymatic treatments (hydrolysis with xylanase or beta-glucanase or a mixture of these two enzymes) were performed, and their effect on sterol content, extractability of sterols and the characteristic steryl conjugates of cereals (steryl ferulates, steryl glycosides, and acylated steryl glycosides) were studied. Mechanical and enzymatic treatments increased the apparent sterol content, whereas aqueous processing without enzymes hindered the availability of total sterols, especially from rye bran. Changes were also seen in the amounts of steryl conjugates caused by the enzymatic treatments. On the basis of the results of this study, it can be speculated that a combination of fine particle size and enzymatic processing results in optimal sterol availability in cereal processing.