The present paper describes an efficient single step chromatographic method for purification of C-Phycocyanin from three cyanobacterial species, i.e., Spirulina sp. (freshwater), Phormidium sp. (marine water) and Lyngbya sp. (marine water). C-Phycocyanin from these cyanobacterial species was purified to homogeneity and some of their properties were investigated. The purification involves a multistep treatment of the crude extract by fractional precipitation with ammonium sulfate, followed by ion-exchange chromatography on DEAE-Sepharose CL-6B column. Pure C-Phycocyanin was finally obtained from Spirulina, Phormidium, and Lyngbya spp. with purity ratio (A620/A280) 4.42, 4.43, and 4.59, respectively, further the purity and homogeneity were confirmed by native and SDS-PAGE. The estimated molecular weights of purified C-PC from Spirulina, Phormidium, and Lyngbya spp. were 112, 131, and 81 kDa, respectively. SDS-PAGE of pure C-Phycocyanin yielded two bands corresponding to alpha and beta subunits. The results of SDS-PAGE demonstrate the same molecular weight of beta subunits (24.4 kDa) for all the three cyanobacterial species, whereas the molecular weight of the alpha subunit is different for all (17 kDa Spirulina sp., 19.1 kDa Phormidium sp., 15.2 kDa Lyngbya sp.). Thus, the C-Phycocyanin was characterized as (alphabeta)3 for Spirulina and Phormidium spp., while as (alphabeta)2 for Lyngbya sp.