Detection of monocytic differentiation in myeloid neoplasms by immunohistochemical analysis is challenging owing to a lack of sensitive and/or specific antibodies. We tested the usefulness of immunohistochemical analysis for CD14, an antigen commonly detected by flow cytometry, and Krüppel-like factor 4 (KLF4), a potentially novel marker of monocytic differentiation, in a series of myeloid leukemias, including 53 acute myeloid leukemias with monocytic differentiation. These findings were compared with immunohistochemical findings for CD68 (KP-1), CD34, and CD163 and were also correlated with flow cytometric and enzyme cytochemical results. CD163 and CD14 are the most specific markers of monocytic differentiation, followed by KLF4. CD68, in contrast, is the most sensitive monocytic marker, and KLF4 is also significantly more sensitive than CD14 and CD163. These studies show that KLF4 is another marker of monocytic differentiation and that the combination of CD14 and CD163 can increase the diagnostic sensitivity for monocytic neoplasms.