Chemokines in biological sample are frequently found at very low level. Further, concentrations of chemokines which were measured in the same tissue detected with different methods often differ in literature. Therefore, RANTES concentrations were quantified by application of sandwich enzyme-linked immunosorbent assay (ELISA) and Western blot analyses. The sensitivity of ELISA was found to be much higher than that of Western blot. Additionally, the detection time differed considerably as well. For biological and biochemical characterization of chemokines it is essential to implement the optimal purification and detection techniques. With an understanding of the technical procedures and some pitfalls, chemokine detection can be applied more reliably.