Abstract: Brucellosis is an extremely important disease affecting the health of human and a number of animal species (including buffalo) around the world and Brucellae sp. is the causative agent of brucellosis. As an important innate immunity molecule, CD14, involved in protecting a host against invading microorganism including Brucella. To analyze the expression of CD14 and provide new alternatives for elucidating the molecular mechanism of Brucella infection in bufflo, real-time quantitative Reverse Transcription PCR (RT-PCR) and western blot were performed to assess the mRNA level and protein levels of CD14 in heart, liver, spleen, lung, kidney and intestine. The results indicated that CD14 mRNA and protein are ubiquitously present in all buffalo tissues examined with relatively high levels in lung and relatively low levels in intestine. These data provide important anatomical information for studying the role of CD14 played in Brucella infection in the different tissue of buffalo.